Immunotherapy of cancer

ABSTRACT

Immunogenic modulators and compositions comprising oligonucleotide agents capable of inhibiting suppression of immune response by reducing expression of one or more gene involved with an immune suppression mechanism.

CROSS REFERENCE

This application is a national stage filing under 35 U.S.C. § 371 of International Application No. PCT/US2014/068244, filed Dec. 2, 2014, which was published under PCT Article 21(2) in English and claims priority to U.S. Provisional Application No. 61/910,728, filed Dec. 2, 2013, each of which is herein incorporated by reference in its entirety.

TECHNICAL FIELD

The present invention relates to immunogenic compositions, method of making immunogenic compositions, and methods of using immunogenic compositions for the treatment of cell proliferative disorders or infectious disease, including, for example, cancer and autoimmune disorders.

More particularly, the invention provides cells that are treated with oligonucleotides specifically designed to modulate expression of target genes involved in tumor immune resistance mechanisms.

BACKGROUND

Immunotherapy is the “treatment of disease by inducing, enhancing, or suppressing an immune response”. Immunotherapies designed to elicit or amplify an immune response are activation immunotherapies, while immunotherapies that reduce or suppress immune response are classified as suppression immunotherapies.

Immunotherapy of cancer has become increasingly important in clinical practice over recent decades. The primary approach in today's standard of care is passive immunotherapy through the use of recombinant monoclonal antibodies (mAbs). MAbs act through a mechanisms relevant to the body's own humoral immune response, by binding to key antigens involved in the tumor development and causing moderate forms of cell-mediated immunity, such as antibody-dependent cell-mediated cytotoxicity (ADCC).

Another group of emerging immunotherapeutic approaches is based on the administration of cells capable of destroying tumor cells. The administered cells may be the patient's own tumor-infiltrating lymphocytes (TIL), isolated and expanded ex-vivo. In some cases, TIL are capable of recognizing a variety of tumor associated antigens (TAA), while in other cases TIL can be reactivated and expanded in vitro to recognize specific antigens. The TIL-based therapeutic approaches are commonly referred to as “adoptive cell transfer” (ACT).

Further developments of ACT involve genetic modifications of T-cells to express receptors that recognize specific tumor-associated antigens (TAA). Such modifications may induce the expression of a specific T-cell receptor (TCR) or of a chimeric antigen receptor (CAR) consisting of TAA-specific antibody fused to CD3/co-stimulatory molecule transmembrane and cytoplasmic domains.

The ACT methods may also be considered as passive immunotherapeutic approaches in that they act directly on the tumor cells without invoking an extended immune response. However, unlike mAbs, ACT agents are capable of fully destroying the tumor cells, as opposed to the blockade of selected receptors and moderate cellular responses such as ADCC.

There is ongoing development of numerous methods of active immunotherapy, which restore the ability of body's own immune system to generate antitumor response. Active immunotherapeutic agents are often called therapeutic cancer vaccines, or just cancer vaccines. Many cancer vaccines are currently in clinical trials, and sipuleucell-T has recently become the first such vaccine approved by the United States FDA.

There are several classes of cancer vaccines using different antigens and different mechanisms of generating cell-mediated immune response. One class of vaccines is based on peptide fragments of antigens selectively expressed by tumor cells. The peptides are administered alone or in combination with immune-stimulatory agents, which may include adjuvants and cytokines, such as granulocyte-macrophage colony-stimulating factor (GM-CSF).

Another class of cancer vaccines is based on modified (e.g. sub-lethally irradiated) tumor cells used as antigens, also in combinations with immuno-stimulatory agents. Vaccines of this type currently in clinical trials are based both on autologous (e.g. OncoVAX, LipoNova) and allogeneic (e.g. Canvaxin, Onyvax-P, GVAX) tumor cell lines.

Yet another class of cancer vaccines uses dendritic cells. By their nature, dendritic cells (DC) are “professional” antigen-presenting cells capable of generating of a strong antigen-dependent cell-mediated immune response and eliciting therapeutic T-cells in vivo. DC-based cancer vaccines usually comprise DCs isolated from patients or generated ex vivo by culturing patient's hematopoietic progenitor cells or monocytes. DCs are further loaded with tumor antigens and sometimes combined with immune-stimulating agents, such as GM-CSF. A large number of DC-vaccines are now in clinical trials, and the first FDA-approved vaccine sipuleucell-T is based on DC.

Mechanisms of Immunosuppression and Therapeutic Approaches to its Mitigation

One of the key physiologic functions of the immune system is to recognize and eliminate neoplastic cells, therefore an essential part of any tumor progression is the development of immune resistance mechanisms. Once developed, these mechanisms not only prevent the natural immune system from effecting the tumor growth, but also limit the efficacy of any immunotherapeutic approaches to cancer. An important immune resistance mechanism involves immune-inhibitory pathways, sometimes referred to as immune checkpoints. The immune-inhibitory pathways play particularly important role in the interaction between tumor cells and CD8+ cytotoxic T-lymphocytes, including ACT therapeutic agents. Among important immune checkpoints are inhibitory receptors expressed on the T-cell surface, such as CTLA-4, PD1 and LAGS, among others.

The importance of the attenuation of immune checkpoints has been recognized by the scientific and medical community. One way to mitigate immunosuppression is to block the immune checkpoints by specially designed agents. The CTLA-4-blocking-antibody, ipilimumab, has recently been approved by the FDA. Several molecules blocking PD1 are currently in clinical development.

Immunosuppression mechanisms also negatively affect the function of dendritic cells and, as a consequence, the efficacy of DC-based cancer vaccines. Immunosuppressive mechanisms can inhibit the ability of DC to present tumor antigens through the MHC class I pathway and to prime naïve CD8+ T-cells for antitumor immunity. Among the important molecules responsible for the immunosuppression mechanisms in DC are ubiquitin ligase A20 and the broadly immune-suppressive protein SOCS1.

The efficacy of immunotherapeutic approaches to cancer can be augmented by combining them with inhibitors of immune checkpoints. Numerous ongoing preclinical and clinical studies are exploring potential synergies between cancer vaccines and other immunotherapeutic agents and checkpoint blocking agents, for example, ipilimumab. Such combination approaches have the potential to result in significantly improved clinical outcomes.

However, there are a number of drawbacks of using cancer immunotherapeutic agents in combination with checkpoint inhibitors. For example, immune checkpoint blockade can lead to the breaking of immune self-tolerance, thereby inducing a novel syndrome of autoimmune/auto-inflammatory side effects, designated “immune related adverse events,” mainly including rash, colitis, hepatitis and endocrinopathies (Corsello, et al. J. Clin. Endocrinol. Metab., 2013, 98:1361).

Reported toxicity profiles of checkpoint inhibitors are different than the toxicity profiles reported for other classes of oncologic agents. Those involve inflammatory events in multiple organ systems, including skin, gastrointestinal, endocrine, pulmonary, hepatic, ocular, and nervous system. (Hodi, 2013, Annals of Oncology, 24: Suppl, i7).

In view of the above, there is a need for new cancer therapeutic agents that can be used in combination with checkpoint inhibitors as well as other classes of oncolytic agents without risk of adverse inflammatory events in multiple organ systems previously reported for checkpoint inhibitors. The immunotherapeutic cells of the invention, prepared by treating cells with a combination oligonucleotide agents targeting genes associated with tumor or infections disease resistance mechanisms, as well as methods of producing such therapeutic cells and methods of treating disease with the produced therapeutic cells, satisfy this long felt need.

SUMMARY OF EMBODIMENTS OF THE INVENTION

The efficacy of immunotherapeutic approaches to cell proliferation disorders and infectious diseases can be augmented by combining them with inhibitors of immune checkpoints. Numerous synergies between cancer vaccines and other immunotherapeutic agents and checkpoint blocking agents provide opportunities for combination approaches that may significantly improve clinical outcomes for example, in proliferative cell disorders and immune diseases.

Various embodiments of the inventions disclosed herein include compositions comprising therapeutic cells obtained by treating cells ex vivo with oligonucleotides to modulate expression of target genes involved in immune suppression mechanisms. The oligonucleotide agent may be an antisense oliogonucleotide (ASO), including locked nucleic acids (LNAs), methoxyethyl gapmers, and the like, or an siRNA, miRNA, miRNA-inhibitor, morpholino, PNA, and the like. The oligonucleotide is preferably a self-delivered (sd) RNAi agent. The oligonucleotides may be chemically modified, for example, including at least one 2-O-methyl modification, 2′-Fluro modification, and/or phosphorothioate modification. The oligonucleotides may include one or more hydrophobic modification, for example, one or more sterol, cholesterol, vitamin D, Naphtyl, isobutyl, benzyl, indol, tryptophane, or phenyl hydrophobic modification. The oligonucleotide may be a hydrophobically-modified siRNA-antisense hybrid. The oligonucleotides may be used in combination with transmembrane delivery systems, such as delivery systems comprising lipids.

In an embodiment, the cells are obtained and/or derived from a cancer or infectious disease patient, and may be, for example, tumor infiltrating lymphocytes (TIL) and/or T-cells, antigen presenting cells such as dendritic cells, natural killer cells, induced-pluripotent stem cells, stem central memory T-cells, and the like. The T-cells and NK-cells are preferably genetically engineered to express high-affinity T-Cell receptors (TCR) and/or chimeric antibody or antibody-fragment—T-Cell receptors (CAR). In an embodiment, the chimeric antibody/antibody fragment is preferably capable of binding to antigens expressed on tumor cells. Immune cells may be engineered by transfection with plasmid, viral delivery vehicles, or mRNAs.

In an embodiment, the chimeric antibody or fragment is capable of binding CD19 receptors of B-cells and/or binding to antigens expressed on tumors, such as melanoma tumors. Such melanoma-expressed antigens include, for example, GD2, GD3, HMW-MAA, VEGF-R2, and the like.

Target genes identified herein for modification include: cytotoxic T-cell antigen 4 (CTLA4), programmed cell death protein 1 (PD1), tumor growth factor receptor beta (TGFR-beta), LAG3, TIM3, and adenosine A2a receptor; anti-apoptotic genes including, but not limited to: BAX, BAC, Casp8, and P53; A20 ubiquitine ligase (TNFAIP3, SOCS1 (suppressor of cytokine signaling), IDO (indolamine-2,3-dioxygenase; tryptophan-degrading enzyme), PD-L1 (CD274)(surface receptor, binder to PD1 on Tcells), Notch ligand Delta1 (DLL1), Jagged 1, Jagged 2, FasL (pro-apoptotic surface molecule), CCL17, CCL22 (secreted chemokines that attract Treg cells), IL10 receptor (IL10RA), p38 (MAPK14), STAT3, TNFSF4 (OX40L), MicroRNA miR-155, miR-146a, anti-apoptotic genes including but not limited to BAX, BAC, Casp8 and P53, and the like genes, and combinations thereof. Representative target sequences are listed in Table 1.

The engineered therapeutic cells are treated with RNAi agents designed to inhibit expression of one or more of the targeted genes. The RNAi agent may comprise a guide sequence that hybridizes to a target gene and inhibits expression of the target gene through an RNA interference mechanism, where the target region is selected from the group listed in Table 1. The RNA agent can be chemically modified, and preferably includes at least one 2′-O-methyl, 2′-O-Fluoro, and/or phosphorothioate modification, as well as at least one hydrophobic modification such as cholesterol, and the like.

The immunogenic compositions described herein are useful for the treatment of proliferative disorders, including cancers, and/or infectious disease and are produced by the ex-vivo treatment of cells with oligonucleotides to modulate the expression of target genes involved in tumor immune resistance mechanisms. The ex vivo treatment of cells includes administering to the cells an oligonucleotide capable of targeting and inhibiting expression of a gene involved in a tumor suppressor mechanism, such as the genes listed in Table 1. The oligonucleotide can be used in combination with a transmembrane delivery system that may comprise one or more of: lipid(s) and vector, such as a viral vector.

The invention includes a method of treating a cell proliferative disorder or infectious disease by administering to a subject in need thereof, an immunogenic composition comprising cells that have been treated with one or more oligonucleotide to modulate the expression of one or more target gene involved in tumor immune resistance mechanisms, for example, one or more of the target genes of Table 1.

The invention preferably includes immunogenic cells treated with a plurality of oligonucleotide agents targeting a combination of target genes described herein. The combination may target a plurality of suppressor receptor genes, cytokine receptor genes, regulatory genes, and/or apoptotic factors in order to inhibit tumor immune resistance mechanisms.

The present invention is directed to novel immunotherapeutic cells, methods of generating the immunotherapeutic cells, and therapeutic methods employing such cells.

A new method of immune checkpoint inhibition is described herein, applicable to a broad variety of cell-based immunotherapies, including, but not limited to adaptive cell transfer, for example, based on TIL, TCR, CAR, and other cell types, as well as dendritic cell-based cancer vaccines. Self-deliverable RNAi technology provides efficient transfection of short oligonucleotides in any cell type, including immune cells, providing increased efficacy of immunotherapeutic treatments. In addition, the activated immune cells can be protected by preventing apoptosis via inhibition of key activators of the apoptotic pathway, such as BAC, BAX, Casp8, and P53, among others.

The activated immune cells modified by oligonucleotide transfer for a single therapeutic agent for administration to a subject, providing a number of advantages as compared to separately administered combinations of vaccines and immunotherapeutics and separately administered checkpoint inhibitors. These advantages include lack of side effects associated with the checkpoint inhibitors in a single therapeutic agent (activated immune cells modified by oligonucleotides targeting immune resistance genes).

The claimed immunotherapeutic cells, method of producing immunotherapeutic cells by introduction of oligonucleotide molecules targeting immune resistance pathways, and methods of treating proliferative and infectious disease, improves upon any known immunotherapeutic cells and methods of producing immunotherapeutic cells because it provides:

-   -   1) a single therapeutic composition providing a combination of         checkpoint inhibitors and other immune resistance mechanism         inhibitors;     -   2) with reduced toxicity; and     -   3) increased efficacy as compared with other compositions.

BRIEF DESCRIPTION OF THE FIGURES

The foregoing features of embodiments will be more readily understood by reference to the following detailed description, taken with reference to the accompanying drawings, in which:

FIG. 1 is a schematic diagram showing the structure of an sdRNA molecule.

FIG. 2 is a graph showing sdRNA-induced silencing of GAPDH and MAP4K4 in HeLa cells.

FIG. 3 is a graph showing sdRNA-induced knock-down of multiple targets using sdRNA agents directed to three genes in NK-92 cells.

FIG. 4 is a graph showing the knock-down of gene expression in Human Primary T cells by sdRNA agents targeting TP53 and MAP4K4.

FIG. 5 is a graph showing sdRNA-induced knock-down of CTLA4 and PD1 in Human Primary T cells.

FIG. 6 is a graph showing the reduction of PDCD1 and CTLA-4 surface expression by sdRNA in Human Primary T cells.

FIG. 7 is a graph showing MAP4K4-cy3 sdRNA delivery into T and B cells in human PBMCs.

DETAILED DESCRIPTION OF SPECIFIC EMBODIMENTS

The invention is defined by the claims, and includes oligonucleotides specifically designed and selected to reduce and/or inhibit expression of suppressors of immune resistance (inhibitory oligonucleotides), compositions comprising cells modified by treatment with such inhibitory oligonucleotides, methods of making such compositions, and methods of using the compositions to treat proliferation and/or infectious diseases. In particular, cells are treated with a combination of oligonucleotide agents, each agent particularly designed to interfere with and reduce the activity of a targeted immune suppressor.

Preferably, the combination of oligonucleotide agents targets multiple immune suppressor genes selected from checkpoint inhibitor genes such as CTLA4, PD-1/PD-1L, BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3, B7-H4 receptors, and TGF beta type 2 receptor; cytokine receptors that inactivate immune cells, such as TGF-beta receptor A and IL-10 receptor; regulatory genes/transcription factors modulating cytokine production by immune cells, such as STAT-3 and P38, miR-155, miR-146a; and apoptotic factors involved in cascades leading to cell death, such as p53 and Cacp8.

Most preferably the oligonucleotide agent is a self-deliverable RNAi agent, which is a hydrophobically modified siRNA-antisense hybrid molecule, comprising a double-stranded region of about 13-22 base pairs, with or without a 3′-overhang on each of the sense and antisense strands, and a 3′ single-stranded tail on the antisense strand of about 2-9 nucleotides. The oligonucleotide contains at least one 2′-O-Methyl modification, at least one 2′-O-Fluoro modification, and at least one phosphorothioate modification, as well as at least one hydrophobic modification selected from sterol, cholesterol, vitamin D, napthyl, isobutyl, benzyl, indol, tryptophane, phenyl, and the like hydrophobic modifiers (see FIG. 1). The oligonucleotide may contain a plurality of such modifications.

Definitions

As used in this description and the accompanying claims, the following terms shall have the meanings indicated, unless the context requires otherwise:

Proliferative disease, as used herein, includes diseases and disorders characterized by excessive proliferation of cells and turnover of cellular matrix, including cancer, atherlorosclerosis, rheumatoid arthritis, psoriasis, idiopathic pulmonary fibrosis, scleroderma, cirrhosis of the liver, and the like. Cancers include but are not limited to, one or more of: small cell lung cancer, colon cancer, breast cancer, lung cancer, prostate cancer, ovarian cancer, pancreatic cancer, melanoma, hematological malignancy such as chronic myeloid leukemia, and the like cancers where immunotherapeutic intervention to suppress tumor related immune resistance is needed.

Immune target genes can be grouped into at least four general categories: (1) checkpoint inhibitors; (2) cytokine receptors that inactivate immune cells, (3) anti-apoptotic genes; and (4) regulator genes, for example, transcription factors.

Immune Checkpoint inhibitors (ICI), as used herein, include immunotherapeutic agents that bind to certain checkpoint proteins, such as cytotoxic T lymphocyte antigen-4 (CTLA-4) and programmed death-1 (PD-1) and its ligand PD-L1 to block and disable inhibitory proteins that prevent the immune system from attacking diseased cells such as cancer cells, liberating tumor-specific T cells to exert their effector function against tumor cells.

Tumor related immune resistance genes, as used herein, include genes involved in checkpoint inhibition of immune response, such as CTLA-4 and PD-1/PD-L1; TGF-beta, LAG3, Tim3, adenosine A2a receptor;

Regulator genes, as used herein, include transcription factors and the like that modulate cytokine production by immune cells, and include p38, STAT3, microRNAs miR-155, miR-146a;

Anti-apoptotic genes, as used herein, include BAX, BAC, Casp8, P53 and the like; and combinations thereof.

Infectious diseases, as used herein, include, but are not limited to, diseases caused by pathogenic microorganisms, including, but not limited to, one or more of bacteria, viruses, parasites, or fungi, where immunotherapeutic intervention to suppress pathogen related immune resistance and/or overactive immune response.

Immunogenic composition, as used herein, includes cells treated with one or more oligonucleotide agent, wherein the cells comprise T-cells. The T-cells may be genetically engineered, for example, to express high affinity T-cell receptors (TCR), chimeric antibody—T-cell receptors (CAR), where the chimeric antibody fragments are capable of binding to CD19 receptors of B-cells and/or to antigens expressed on tumor cells. In one embodiment, the chimeric antibody fragments bind antigens expressed on melanoma tumors, selected from GD2, GD3, HMW-MAA, and VEGF-R2.

Immunogenic compositions described herein include cells comprising antigen-presenting cells, dendritic cells, engineered T-cells, natural killer cells, stem cells, including induced pluripotent stem cells, and stem central memory T-cells. The treated cell also comprises one or a plurality of oligonucleotide agents, preferably sdRNAi agents specifically targeting a gene involved in an immune suppression mechanism, where the oligonucleotide agent inhibits expression of said target gene.

In one embodiment, the target gene is selected from A20 ubiquitin ligase such as TNFAIP3, SOCS1 (suppressor of cytokine signaling), Tyro3/Ax1/Mer (suppressors of TLR signaling), IDO (indolamine-2,3-dioxygenase, tryptophan-degrading enzyme), PD-L1/CD274 (surface receptor, binds PD1 on T-cells), Notch ligand Delta (DLL1), Jagged 1, Jagged 2, FasL (pro-apoptotic surface molecule), CCL17, CCL22 (secreted chemokines that attract Treg cells), IL-10 receptor (IL10Ra), p38 (MAPK14), STAT3, TNFSF4 (OX40L), microRNA miR-155, miR-146a, anti-apoptotic genes, including but not limited to BAX, BAC, Casp8, and P53; and combinations thereof.

Particularly preferred target genes are those shown in Table 1.

Ex-vivo treatment, as used herein, includes cells treated with oligonucleotide agents that modulate expression of target genes involved in immune suppression mechanisms. The oligonucleotide agent may be an antisense oligonucleotide, including, for example, locked nucleotide analogs, methyoxyethyl gapmers, cyclo-ethyl-B nucleic acids, siRNAs, miRNAs, miRNA inhibitors, morpholinos, PNAs, and the like. Preferably, the oligonucleotide agent is an sdRNAi agent targeting a gene involved in an immune suppression mechanism. The cells treated in vitro by the oligonucleotide agent may be immune cells expanded in vitro, and can be cells obtained from a subject having a proliferative or infectious disease. Alternatively, the cells or tissue may be treated in vivo, for example by in situ injection and/or intravenous injection.

Oligonucleotide or oligonucleotide agent, as used herein, refers to a molecule containing a plurality of “nucleotides” including deoxyribonucleotides, ribonucleotides, or modified nucleotides, and polymers thereof in single- or double-stranded form. The term encompasses nucleotides containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides. Examples of such analogs include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs).

Nucleotide, as used herein to include those with natural bases (standard), and modified bases well known in the art. The nucleotides can be unmodified or modified at the sugar, phosphate and/or base moiety, (also referred to interchangeably as nucleotide analogs, modified nucleotides, non-natural nucleotides, non-standard nucleotides and other; see, for example, PCT Publications No. WO 92/07065 and WO 93/15187. Non-limiting examples of base modifications that can be introduced into nucleic acid molecules include, hypoxanthine, purine, pyridin-4-one, pyridin-2-one, phenyl, pseudouracil, 2,4,6-trimethoxy benzene, 3-methyl uracil, dihydrouridine, naphthyl, aminophenyl, 5-alkylcytidines (e.g., 5-methylcytidine), 5-alkyluridines (e.g., ribothymidine), 5-halouridine (e.g., 5-bromouridine) or 6-azapyrimidines or 6-alkylpyrimidines (e.g. 6-methyluridine and pseudouridine), propyne, and others. The phrase “modified bases” includes nucleotide bases other than adenine, guanine, cytosine, and uracil, modified for example, at the 1′ position or their equivalents.

As used herein, the term “deoxyribonucleotide” encompasses natural and synthetic, unmodified and modified deoxyribonucleotides. Modifications include changes to the sugar moiety, to the base moiety and/or to the linkages between deoxyribonucleotide in the oligonucleotide.

As used herein, the term “RNA” defines a molecule comprising at least one ribonucleotide residue. The term “ribonucleotide” defines a nucleotide with a hydroxyl group at the 2′ position of a □-D-ribofuranose moiety. The term RNA includes double-stranded RNA, single-stranded RNA, isolated RNA such as partially purified RNA, essentially pure RNA, synthetic RNA, recombinantly produced RNA, as well as altered RNA that differs from naturally occurring RNA by the addition, deletion, substitution and/or alteration of one or more nucleotides. Nucleotides of the RNA molecules described herein may also comprise non-standard nucleotides, such as non-naturally occurring nucleotides or chemically synthesized nucleotides or deoxynucleotides. These altered RNAs can be referred to as analogs or analogs of naturally-occurring RNA.

As used herein, “modified nucleotide” refers to a nucleotide that has one or more modifications to the nucleoside, the nucleobase, pentose ring, or phosphate group. For example, modified nucleotides exclude ribonucleotides containing adenosine monophosphate, guanosine monophosphate, uridine monophosphate, and cytidine monophosphate and deoxyribonucleotides containing deoxyadenosine monophosphate, deoxyguanosine monophosphate, deoxythymidine monophosphate, and deoxycytidine monophosphate. Modifications include those naturally-occurring that result from modification by enzymes that modify nucleotides, such as methyltransferases.

Modified nucleotides also include synthetic or non-naturally occurring nucleotides. Synthetic or non-naturally occurring modifications in nucleotides include those with 2′ modifications, e.g., 2′-O-methyl, 2′-methoxyethoxy, 2′-fluoro, 2′-allyl, 2′-O-[2-(methylamino)-2-oxoethyl], 4′-thio, 4′-CH2-O-2′-bridge, 4′-(CH2) 2-O-2′-bridge, 2′-LNA, and 2′-O—(N-methylcarbamate) or those comprising base analogs. In connection with 2′-modified nucleotides as described for the present disclosure, by “amino” is meant 2′-NH2 or 2′-O—NH2, which can be modified or unmodified. Such modified groups are described, for example, in U.S. Pat. Nos. 5,672,695 and 6,248,878.

As used herein, “microRNA” or “miRNA” refers to a nucleic acid that forms a single-stranded RNA, which single-stranded RNA has the ability to alter the expression (reduce or inhibit expression; modulate expression; directly or indirectly enhance expression) of a gene or target gene when the miRNA is expressed in the same cell as the gene or target gene. In one embodiment, a miRNA refers to a nucleic acid that has substantial or complete identity to a target gene and forms a single-stranded miRNA. In some embodiments miRNA may be in the form of pre-miRNA, wherein the pre-miRNA is double-stranded RNA. The sequence of the miRNA can correspond to the full length target gene, or a subsequence thereof. Typically, the miRNA is at least about 15-50 nucleotides in length (e.g., each sequence of the single-stranded miRNA is 15-50 nucleotides in length, and the double stranded pre-miRNA is about 15-50 base pairs in length). In some embodiments the miRNA is 20-30 base nucleotides. In some embodiments the miRNA is 20-25 nucleotides in length. In some embodiments the miRNA is 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.

Target gene, as used herein, includes genes known or identified as modulating the expression of a gene involved in an immune resistance mechanism, and can be one of several groups of genes, such as suppressor receptors, for example, CTLA4 and PD1; cytokine receptors that inactivate immune cells, for example, TGF-beta receptor, LAG3, Tim3, adenosine A2a receptor, and IL10 receptor; regulatory genes for example, STAT3, p38, mir155 and mir146a; and apoptosis factors involved in cascades leading to cell death, for example, P53, Casp8, BAX, BAC, and combinations thereof. See also preferred target genes listed in Table 1.

As used herein, small interfering RNA (siRNA), sometimes known as short interfering RNA or silencing RNA, defines a group of double-stranded RNA molecules, comprising sense and antisense RNA strands, each generally of about 1022 nucletides in length, optionally including a 3′ overhang of 1-3 nucleotides. siRNA is active in the RNA interference (RNAi) pathway, and interferes with expression of specific target genes with complementary nucleotide sequences.

As used herein, sdRNA refers to “self-deliverable” RNAi agents, that are formed as an asymmetric double-stranded RNA-antisense oligonucleotide hybrid. The double stranded RNA includes a guide (sense) strand of about 19-25 nucleotides and a passenger (antisense) strand of about 10-19 nucleotides with a duplex formation that results in a single-stranded phosphorothiolated tail of about 5-9 nucleotides.

The RNA sequences may be modified with stabilizing and hydrophobic modifications such as sterols, for example, cholesterol, vitamin D, naphtyl, isobutyl, benzyl, indol, tryptophane, and phenyl, which confer stability and efficient cellular uptake in the absence of any transfection reagent or formulation. Immune response assays testing for IFN-induced proteins indicate sdRNAs produce a reduced immunostimulatory profile as compared other RNAi agents. See, for example, Byrne et al., December 2013, J. Ocular Pharmacology and Therapeutics, 29(10): 855-864.

Cell-Based Immunotherapeutics

In general, cells are obtained from subjects with proliferative disease such as cancer, or an infectious disease such as viral infection. The obtained cells are treated directly as obtained or may be expanded in cell culture prior to treatment with oligonucleotides. The cells may also be genetically modified to express receptors that recognize specific antigens expressed on the tumor cell surface (CAR) or intracellular tumor antigens presented on MHC class I (TCR).

Oligonucleotide Agents

Antisense Oligonucleotides

Small interfering RNA (siRNA), sometimes known as short interfering RNA or silencing RNA, is a double stranded RNA molecule, generally 19-25 base pairs in length. siRNA is used in RNA interference (RNAi), where it interferes with expression of specific genes with complementary nucleotide sequences.

Double stranded DNA (dsRNA) can be generally used to define any molecule comprising a pair of complementary strands of RNA, generally a sense (passenger) and antisense (guide) strands, and may include single-stranded overhang regions. The term dsRNA, contrasted with siRNA, generally refers to a precursor molecule that includes the sequence of an siRNA molecule which is released from the larger dsRNA molecule by the action of cleavage enzyme systems, including Dicer.

sdRNA (self-deliverable) are a new class of covalently modified RNAi compounds that do not require a delivery vehicle to enter cells and have improved pharmacology compared to traditional siRNAs. “Self-deliverable RNA” or sdRNA is a hydrophobically modified RNA interfering-antisense hybrid, demonstrated to be highly efficacious in vitro in primary cells and in vivo upon local administration. Robust uptake and/or silencing without toxicity has been demonstrated in several tissues including dermal, muscle, tumors, alveolar macrophages, spinal cord, and retina cells and tissues. In dermal layer and retina, intradermal and intra-vitreal injection of sdRNA at mg doses induced potent and long lasting silencing.

While sdRNA is a superior functional genomics tool, enabling RNAi in primary cells and in vivo, it has a relatively low hit rate as compared to conventional siRNAs. While the need to screen large number of sequences per gene is not a limiting factor for therapeutic applications, it severely limits the applicability of sdRNA technology to functional genomics, where cost effective compound selection against thousands of genes is required. To optimize sdRNA structure, chemistry, targeting position, sequence preferences, and the like, a proprietary algorithm has been developed and utilized for sdRNA potency prediction. Availability of sdRNA reagents that are active in all cell types ex vivo and in vivo enables functional genomics and target stratification/validation studies.

Proprietary Algorithm

SdRNA sequences were selected based on a proprietory selection algorithm, designed on the basis of a functional screen of over 500 sdRNA sequences in the luciferase reporter assay of HeLa cells. Regression analysis of was used to establish a correlation between the frequency of occurrence of specific nucleotide and modification at any specific position in sdRNA duplex and its functionality in gene suppression assay. This algorithm allows prediction of functional sdRNA sequences, defined as having over 70% knockdown μM concentration, with a probability over 40%.

Table 1 shows predictive gene targets identified using the proprietary algorithm and useful in the cellular immunotherapeutic compositions and methods described herein.

Delivery of RNAi Agents

BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; Applic BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; ation of RNAi technology to functional genomics studies in prim BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; ary cells and in vivo is limited by requirements to formulate siRNAs into lipids or use of other cell delivery techniques. To circumvent delivery problems, the self-deliverable RNAi technology provides a method of directly transfecting cells with the RNAi agent, without the need for additional formulations or techniques. The ability to transfect hard-to-transfect cell lines, high in vivo activity, and simplicity of use, are characteristics of the compositions and methods that present significant functional advantages over traditional siRNA-based techniques. The sdRNAi technology allows direct delivery of chemically synthesized compounds to a wide range of primary cells and tissues, both ex-vivo and in vivo.

To enable BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; self-delivery, traditional siRNA molecules require a substantial reduction in size and the introduction of extensive chemical modifications which are not well tolerated by RNAi machinery, resulting in extremely low probability of finding active molecules (low hit rate). In contrast, the sdRNA technology allows efficient RNAi delivery to primary cells and tissues in vitro and in vivo, with demonstrated silencing efficiency in humans.

The general structure of sdRNA molecules is shown in FIG. 1. sdRNA are formed as hydrophobically-modified siRNA-antisense oligonucleotide hybrid structures, and are disclosed, for example in Byrne et al., Dec. 2013, J. Ocular Pharmacology and Therapeutics, 29(10): 855-864.

Oligonucleotide Modifications: 2′-O-Methyl, 2′-O-Fluro, Phosphorothioate

The oligonucleotide agents preferably comprise one or more modification to increase stability and/or effectiveness of the therapeutic agent, and to effect efficient delivery of the oligonucleotide to the cells or tissue to be treated. Such modifications include at least one

BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; 2′-O-methyl modification, at least one 2′-O-Fluro modification, and at least one diphosphorothioate modification. Additionally, the oligonucleotide is modified to include one or more hydrophobic modification selected from sterol, cholesterol, vitamin D, naphtyl, isobutyl, benzyl, indol, tryptophane, and phenyl. The hydrophobic modification is preferably a sterol.

Delivery of Oligonucleotide Agents to Cells

The oligonucleotides may be delivered to the cells in combination with a transmembrane delivery system, preferably comprising lipids, viral vectors, and the like. Most preferably, the oligonucleotide agent is a self-delivery RNAi agent, that does not require any delivery agents.

Combination Therapy

Most preferred for this invention, e.g. particular combinations of elements and/or alternatives for specific needs. This objective is accomplished by determining the appropriate genes to be targeted by the oligonucleotide in order to silence immune suppressor genes and using the proprietary algorithm to select the most appropriate target sequence.

It is preferred that the immunotherapeutic cell be modified to include multiple oligonucleotide agents targeting a variety of genes involved in immune suppression and appropriate for the selected target disease and genes. For example, a preferred immunotherapeutic cell is a T-Cell modified to knock-down both CTLA-4 and PD-1

Additional combinations of oligonucleotides to related genes involved in immune suppression include varied combinations of the selected target sequences of Table 1.

BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; Preferred BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; therapeutic combinations include cells engineered to knock down gene expression of the following target genes:

a) CTLA4 and PD1

b) STAT3 and p38

c) PD1 and BaxPD1, CTLA4, Lag-1, ILM-3, and TP53

d) PD1 and Casp8

e) PD1 and IL10R

The therapeutic compositions described herein are useful to treat a subject suffering from a proliferation disorder or infectious disease. In particular, the immunotherapeutic composition is useful to treat disease characterized by suppression of the subjects immune mechanisms. The sdRNA agents described herein are specifically designed to target genes involved in diseases-associated immune suppression pathways.

Methods of treating a subject comprise administering to a subject in need thereof, an immunogenic composition comprising an sdRNAi agent capable of inhibiting expression of genes involved in immune suppression mechanisms, for example, any of the genes listed in Table 1 or otherwise described herein.

EXAMPLES

The embodiments of the invention described above are intended to be merely exemplary; numerous variations and modifications will be apparent to those skilled in the art. All such variations and modifications are intended to be within the scope of the present invention as defined in any appended claims.

Example 1 Self-Deliverable RNAi Immunotherapeutic Agents

Immunotherapeutic agents described herein were produced by treating cells with particular sdRNA agents designed to target and knock down specific genes involved in immune suppression mechanisms. In particular, the following cells and cell lines have been successfully treated with sdRNA and were shown to knock down at least 70% of targeted gene expression in the specified human cells.

These studies demonstrated utility of these immunogenic agents to suppress expression of target genes in cells normally very resistant to transfection, and suggests the agents are capable of reducing expression of target cells in any cell type.

TABLE 2 Target Cell Type Gene sdRNA target sequence % Knock Down Primary human TP53 (P53) GAGTAGGACATACCAGCTTA >70% 2 uM T-cells (SEQ ID NO: 1001) Primary human MAP4K4 AGAGTTCTGTGGAAGTCTA >70% 2 uM T-cells (SEQ ID NO: 1002) Jurkat T-lymphoma MAP4K4 AGAGTTCTGTGGAAGTCTA 100% 1 uM 72h cells (SEQ ID NO: 1003) NK-92 cells MAP4K4 AGAGTTCTGTGGAAGTCTA  80% 2 uM 72h (SEQ ID NO: 1004) NK-92 cells PPIB ACAGCAAATTCCATCGTGT >75% 2 uM 72h (SEQ ID NO: 1005) NK-92 cells GADPH CTGGTAAAGTGGATATTGTT >90% 2 uM 72h (SEQ ID NO: 1006) HeLa Cells MAP4K4 AGAGTTCTGTGGAAGTCTA >80% 2 uM 72h (SEQ ID NO: 1007)

Example 2 Oligonucleotide Sequences for Inhibiting Expression of Target Genes

A number of human genes were selected as candidate target genes due to involvement in immune suppression mechanisms, including the following genes shown in Table 3:

BAX BAK1 CASP8 (NM_004324) (NM_001188) (NM_001228) ADORA2A CTLA4 LAG3 (NM_000675) (NM_005214) (NM002286) PDCD1 TGFBR1 HAVCR2 (NM_NM005018) (NM-004612) (NM_032782) CCL17 CCL22 DLL2 (NM_002987) (NM_002990) (NM_005618) FASLG CD274 IDO1 (NM_000639) (NM_001267706) (NM_002164) IL10RA JAG1 JAG2 (NM_001558) (NM_000214) (NM_002226) MAPK14 SOCS1 STAT3 (NM_001315) (NM_003745) (NM_003150) TNFA1P3 TNFSF4 TYRO2 (NM_006290) (NM_003326) (NM_006293) TP53 (NM_000546)

Each of the genes listed above was analyzed using a proprietary algorithm to identify preferred sdRNA targeting sequences and target regions for each gene for prevention of immunosuppression of antigen-presenting cells and T-cells. Results are shown in Table 1.

Example 3 Knock-Down of Target Gene (GAPDH) by sdRNA in HeLa Cells

HeLa cells (ATCC CRM-CCL-2) were subcultured 24 hours before transfection and kept log phase. The efficacy of several GAPDH sdRNAs was tested by qRT-PCR, including G13 sdRNA listed in the Table 1.

Solutions of GAPDH, MAP4K4 (positive control) and NTC (non-targeting control) sdRNA with twice the required concentration were prepared in serum-free EMEM medium, by diluting 100 μM oligonucleotides to 0.2-4 μM.

The total volume of medium for each oligo concentration point was calculated as [50 μl/well]×[number of replicates for each serum point]. Oligonucleotides were dispensed into a 96 well plate at 50 μl/well.

Cells were collected for transfection by trypsinization in a 50 ml tube, washed twice with medium containing 10% FBS without antibiotics, spun down at 200×g for 5 minutes at room temperature and resuspended in EMEM medium containing twice the required amount of FBS for the experiment (6%) and without antibiotics. The concentration of the cells was adjusted to 120,000/ml to yield a final concentration of 6,000 cells/50 μl/well. The cells were dispensed at 50 μl/well into the 96-well plate with pre-diluted oligos and placed in the incubator for 48 hours.

Gene Expression Analysis in HeLa Cells Using qRT-PCR

RNA was isolated from transfected HeLa cells using the PureLink™ Pro96 total RNA purification Kit (Ambion, Cat. No. 12173-011A), with Quanta qScript XLT One-Step RT-qPCR ToughMix, ROX (VWR, 89236672). The isolated RNA was analyzed for gene expression using the Human MAP4K4-FAM (Taqman Hs0377405_ml) and Human GAPDH-VIC (Applied Biosystems, Cat. No. 4326317E) gene expression assays.

The incubated plate was spun down and washed once with 100 μl/well PBS and lysed with 60 μl/well buffer provided in the kit. RNA isolation was conducted according to the manufacturer's instructions, and the RNA was eluted with 100 μl RNase-free water, and used undiluted for one-step qRT-PCR.

Dilutions of non-transfected (NT) cells of 1:5 and 1:25 were prepared for the standard curve using RNase-free water. qRT-PCR was performed by dispensing 9 μl/well into a low profile PCR plate and adding 1 μl RNA/well from the earlier prepared RNA samples. After brief centrifugation, the samples were placed in the real-time cycler and amplified using the settings recommended by the manufacturer.

GAPDH gene expression was measured by qPCR, normalized to MAP4K4 and plotted as percent of expression in the presence of non-targeting sdRNA. The results were compared to the normalized according to the standard curve. As shown in FIG. 2, several sdRNA agents targeting GAPDH or MAP4K4 significantly reduced their mRNA levels leading to more than 80-90% knock-down with 1 μM sdRNA. (See FIG. 2).

Example 4 Silencing of Multiple Targets by sdRNA in NK-92 Cells

NK-92 cells were obtained from Conqwest and subjected to one-step RT-PCR analysis without RNA purification using the FastLane Cell Multiplex Kit (Qiagen, Cat. No. 216513). For transfection, NK-92 cells were collected by centrifugation and diluted with RPMI medium containing 4% FBS and IL2 1000 U/ml and adjusted to 1,000,000 cells/ml.

Multiple sdRNA agents targeting MAP4K4, PPIB or GADPH were diluted separately in serum-free RPMI medium to 4 μM and individually aliquoted at 50 μl/well into a 96-well plate. The prepared cells were then added at 50 μl cells/well to the wells with either MAP4K4, PPIB or GAPDH sdRNAs. Cells were incubated for 24, 48, or 72 hours.

At the specified timepoints, the plated transfected cells were washed once with 100 μl/well PBS and once with FCW buffer. After removal of supernatant, cell processing mix of 23.5 μl FCPL and 1.5 μl gDNA wipeout solution was added to each well and incubated for five minutes at room temperature. Lysates were then transferred to PCR strips and heated at 75° C. for five minutes.

To setup qRT-PCR, the lysates were mixed with QuantiTect reagents from the FastLane Cell Multiplex Kit and with primer probe mix for MAP4K4-FAM/GAPDH-VIC or PPIB-FAM/GAPDH-VIC. The following Taqman gene expression assays were used: human MAP4K4-FAM (Taqman, Hs00377405_ml), human PPIB-FAM (Taqman, Hs00168719_ml) and human GAPDH-VIC (Applied Biosystems, cat. No 4326317E).

A volume of 9 μl/well of each reaction mix was dispensed into a low profile PCR plate. One μl lysate per well was added from the previously prepared lysates. The samples were amplified using the settings recommended by the manufacturer.

Results shown in FIG. 3 demonstrate significant silencing of each of the multiple targets, MAP4K4, PPIB, and GADPH by sdRNA agents transfected into NK-92 cells, including greater than 75% inhibition of expression of each target within 24 to 72 hours of incubation.

Example 5 Silencing of TP53 and MAP4K4 by sdRNA in Human Primary T-Cells

Primary human T-cells were obtained from AllCells (CA) and cultured in complete RPMI medium containing 1000 IU/ml IL2. Cells were activated with anti-CD3/CD28 Dynabeads (Gibco, 11131) according to the manufacturer's instructions for at least 4 days prior to the transfection. Cells were collected by brief vortexing to dislodge the beads from cells and separating them using the designated magnet.

sdRNA agents targeting TP53 or MAP4K4 were prepared by separately diluting the sdRNAs to 0.2-4 μM in serum-free RPMI per sample (well) and individually aliquoted at 100 μl/well of 96-well plate. Cells were prepared in RPMI medium containing 4% FBS and IL2 2000 U/ml at 1,000,000 cells/ml and seeded at 100 μl/well into the 96-well plate with pre-diluted sdRNAs.

At the end of the transfection incubation period, the plated transfected cells were washed once with 100 μl/well PBS and processed with FastLane Cell Multiplex Kit reagents essentially as described for the Example 4 and according to the manufacturer's instructions. Taqman gene expression assays were used in the following combinations: human MAP4K4-FAM/GAPDH-VIC or human TP53-FAM (Taqman, Hs01034249 ml)/GAPDH-VIC. A volume of 18 μl/well of each reaction mix was combined with 2 μl lysates per well from the previously prepared lysates. The samples were amplified as before (see Example 4).

Results shown in FIG. 4 demonstrate significant silencing of both MAP4K4 and TP53 by sdRNA agents transfected into T-cells, reaching 70-80% inhibition of gene expression with 1-2 μM sdRNA.

Example 6 Immunotherapeutic Combination of sdRNAs for Treating Melanoma

Melanomas utilize at least two particular pathways to suppress immune function of T-cells, and each involves both PD1 and CTLA4. Melanoma tumors expressing the PD1 ligand, PD1L, can be targeted with T-cells pretreated ex-vivo with sd-RNAi agents specifically designed to target PD1 and interfere with PD1 expression. PD1 is also known as PDCD1, and particular targeting sequences and gene regions identified and predicted to be particularly functional in sdRNA mediated suppression, are shown in Table 1 for PDCD1 (NM_005018) and for CTLA4 (NM005214).

Treatment of melanoma tumors can be effected by providing to melanoma cells T-cells, such as tumor-infiltrating lymphocytes, pretreated ex-vivo with a combination of sdRNAs targeting PD1/PDCD1 and CTLA4, for example, targeting one or more of the twenty target sequences listed for PD1/PDCD1 and/or CTLA4. A combination of sdRNAs targeting PD1/PDCD1 and FASLG (NM_000639) and/or CTLA4, can increase T-cell toxicity in tumors expressing both PD1L and FAS.

In addition to and in combination with anti-CTLA-4 and anti-PD1 sdRNAs, T-cells used for the immunotherapy of melanoma can also be treated with sdRNA targeting other genes implicated in immunosuppression by the tumor. These receptors include, but are not limited to TGF-beta type 1 and 2 receptors, BTLA (binder of herpes virus entry indicator (HVEM) expressed on melanoma cells), and receptors of integrins expressed by myeloid derived suppressor cells (MDSC), such as CD11b, CD18, and CD29.

For tumors whose profile of expressed suppressive proteins is unknown, any combination of sdRNAs targeting PD1/PDCD1 and any one of know suppressing receptors may be helpful to reduce immune suppression and increase therapeutic efficacy.

Example 7 Combination of sdRNAs for Mitigating Immune Cell Suppression

T-cell or dendritic cell suppression may be modulated by various cytokines, such as IL10 and/or TGF beta. Suppressing corresponding receptors in T-cells and dendritic cells may be beneficial for their activity. For example, providing a combination of anti-PD1 with anti-IL10R sdRNAs is expected to mitigate cytokine induced suppression of T-cells and dendritic cells, as compared with anti-PD1 alone.

Example 8 Combination of sdRNAs for Mitigating Immune Cell Suppression

When the mechanism of tumor suppression of immune cells may be not known, use of sdRNA agents to suppress genes involved in apoptosis (programmed cell death), such as p53, Casp8 or other gene activating apoptosis may be beneficial to increase immune cell activity. Combination of an anti-receptor sdRNAs with sdRNAs against pro-apoptotic genes can additionally reduce death of immune cells and thus increase their activity. For example, combination of anti-PD1 with anti-p53 sdRNAs may additionally protect T-cells from suppression by blocking activation of apoptosis.

Example 9 Silencing of CTLA-4 and PDCD1 by sdRNA in Human Primary T-Cells

Primary human T-cells were cultured and activated essentially as described in Example 5. sdRNA agents targeting PDCD1 and CTLA-4 were prepared by separately diluting the sdRNAs to 0.4-4 μM in serum-free RPMI per sample (well) and aliquoted at 100 μl/well of 96-well plate. Cells were prepared in RPMI medium containing 4% FBS and IL2 2000 U/ml at 1,000,000 cells/ml and seeded at 100 μl/well into the 96-well plate with pre-diluted sdRNAs.

72 h later, the transfected cells were washed once with 100 μl/well PBS and processed with FastLane Cell Multiplex Kit reagents essentially as described for the Example 4 and according to the manufacturer's instructions. Taqman gene expression assays were used in the following combinations: human PDCD1-FAM (Taqman, Hs01550088_ml)/GAPDH-VIC or human CTLA4-FAM (Taqman, Hs03044418_ml)/GAPDH-VIC. A volume of 18 μl/well of each reaction mix was combined with 2 μl lysates per well from the previously prepared lysates. The samples were amplified as before (see Example 4).

Results shown in FIG. 5 demonstrate significant silencing of PDCD1 and CTLA-4 by using combined sdRNA agents delivered to T-cells, obtaining greater than 60-70% inhibition of gene expression with 2 μM sdRNA.

Example 10 Reduction of CTLA-4 and PDCD1 Surface Expression by sdRNA in Human Primary T-Cells

Primary human T-cells were cultured and activated essentially as described in Example 5.

sdRNA agents targeting CTLA-4 or PD1 were separately diluted to 5 μM in serum-free RPMI per sample (well) and aliquoted at 250 μl/well to 24-well plates. Cells mixed with magnetic beads were collected and adjusted to 500,000 cells in 250 μl RPMI medium containing 4% FBS and IL2 2000 IU/ml. Cells were seeded at 250 μl/well to the prepared plate containing pre-diluted sdRNAs. 24 hours later FBS was added to the cells to obtain 10% final concentration.

After 72 hours of incubation, the transfected cells were collected, separated from the activation beads using the magnet, as described in Example 5. Cells were washed with PBS, spun down and resuspended in blocking buffer (PBS with 3% BSA) at 200,000 cells/50 μl/sample.

Antibody dilutions were prepared in the blocking buffer. The antibodies were mixed in two combinations: anti-PD1/anti-CD3 (1:100 dilutions for both antibodies) and anti-CTLA4/anti-CD3 (10 μl/106 cells for anti-CTLA4; 1:100 for CD3). The following antibodies were used: rabbit monoclonal [SP7] to CD3 (Abcam, ab16669); mouse monoclonal [BNI3] to CTLA4 (Abcam, ab33320) and mouse monoclonal [NAT105] to PD1 (Abcam, ab52587). Cells were mixed with the diluted antibodies and incubated 30 minutes on ice. Cells were then washed twice with PBS containing 0.2% Tween-20 and 0.1% sodium azide.

Secondary antibodies were diluted in blocking buffer and mixed together resulting in a final dilution 1:500 for anti-mouse Cy5 (Abcam, ab97037) and 1:2000 for anti-rabbit Alexa-488 (Abcam, ab150077). Cells were mixed with the diluted antibodies at 1:1 ratio and incubated 30 minutes on ice. Cells were washed as before, and diluted in 500 μl PBS per tube. The data was acquired immediately on the Attune Acoustic Focusing Cytometer (Applied Biosystems).

As shown in FIG. 6, sdRNA efficiently reduced surface expression of CTLA-4 and PD1 in activated Human Primary T cells.

Example 11 MAP4K4 sdRNA Delivery into CD3- and CD19-Positive Subsets of Human Peripheral Blood Mononuclear Cells (PBMCs)

PBMCs were cultured in complete RPMI supplemented with 1.5% PHA solution and 500 U/ml IL2. For transfection, PBMCs were collected by centrifugation and diluted with RPMI medium containing 4% FBS and IL2 1000 U/ml and seeded to 24-well plate at 500,000 cells/well.

MAP4K4 sdRNA labeled with cy3 was added to the cells at 0.1 μM final concentration. After 72 hours of incubation, the transfected cells were collected, washed with PBS, spun down and diluted in blocking buffer (PBS with 3% BSA) at 200,000 cells/50 μl/sample.

Antibody dilutions were prepared in the blocking buffer as following: 1:100 final dilution anti-CD3 (Abcam, ab16669) and anti-CD19 at 10 μl/1,000,000 cells (Abcam, ab31947). Cells were mixed with the diluted antibodies and incubated 30 min on ice. Cells were then washed twice with PBS containing 0.2% Tween-20 and 0.1% sodium azide.

Secondary antibodies were diluted in the blocking buffer in a final dilution 1:500 for anti-mouse Cy5 (Abcam, ab97037) and 1:2000 for anti-rabbit Alexa-488 (Abcam, ab150077). Cells were mixed with the diluted antibodies at 1:1 ratio and incubated 30 min on ice. Cells were washed as before, and diluted in 500 μl PBS per tube. The data was acquired immediately on the Attune Acoustic Focusing Cytometer (Applied Biosystems).

FIG. 7 shows efficient transfection over 97% of CD3-positive (t cells) and over 98% CD19-positive (B-cells) subsets in Human Peripheral Blood Mononuclear Cells (PBMCs).

TABLE 1 Targeting sequences and gene regions of genes targeted with sdRNAs to prevent immunosuppression of antigen-presenting cells and T-cells. Accession: NM_004324 HUGO gene BAX symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 BAX_NM_004324_human_835 GAATTGCTCAAGTTCATTGA  1 CCTCCACTGCCTCTGGAATTGCTCAAG 21 TTCATTGATGACCCTCTG  2 BAX_NM_004324_human_157 TTCATCCAGGATCGAGCAGG  2 CTTTTGCTTCAGGGTTTCATCCAGGAT 22 CGAGCAGGGCGAATGGGG  3 BAX_NM_004324_human_684 ATCATCAGATGTGGTCTATA  3 TCTCCCCATCTTCAGATCATCAGATGT 23 GGTCTATAATGCGTTTTC  4 BAX_NM_004324_human_412 TACTTTGCCAGCAAACTGGT  4 GTTGTCGCCCTTTTCTACTTTGCCAGCA 24 AACTGGTGCTCAAGGCC  5 BAX_NM_004324_human_538 GGTTGGGTGAGACTCCTCAA  5 ATCCAAGACCAGGGTGGTTGGGTGAG 25 ACTCCTCAAGCCTCCTCAC  6 BAX_NM_004324_human_411 CTACTTTGCCAGCAAACTGG  6 GGTTGTCGCCCTTTTCTACTTTGCCAGC 26 AAACTGGTGCTCAAGGC  7 BAX_NM_004324_human_706 GCGTTTTCCTTACGTGTCTG  7 GATGTGGTCTATAATGCGTTTTCCTTA 27 CGTGTCTGATCAATCCCC  8 BAX_NM_004324_human_716 TACGTGTCTGATCAATCCCC  8 ATAATGCGTTTTCCTTACGTGTCTGATC 28 AATCCCCGATTCATCTA  9 BAX_NM_004324_human_150 TCAGGGTTTCATCCAGGATC  9 AGGGGCCCTTTTGCTTCAGGGTTTCAT 29 CCAGGATCGAGCAGGGCG 10 BAX_NM_004324_human_372_ TGACGGCAACTTCAACTGGG 10 AGCTGACATGTTTTCTGACGGCAACTT 30 CAACTGGGGCCGGGTTGT 11 BAX_NM_004324_human_356 CAGCTGACATGTTTTCTGAC 11 TCTTTTTCCGAGTGGCAGCTGACATGT 31 TTTCTGACGGCAACTTCA 12 BAX_NM_004324_human_357 AGCTGACATGTTTTCTGACG 12 CTTTTTCCGAGTGGCAGCTGACATGTT 32 TTCTGACGGCAACTTCAA 13 BAX_NM_004324_human_776 CACTGTGACCTTGACTTGAT 13 AGTGACCCCTGACCTCACTGTGACCTT 33 GACTTGATTAGTGCCTTC 14 BAX_NM_004324_human_712 TCCTTACGTGTCTGATCAAT 14 GTCTATAATGCGTTTTCCTTACGTGTCT 34 GATCAATCCCCGATTCA 15 BAX_NM_004324_human_465 GATCAGAACCATCATGGGCT 15 CAAGGTGCCGGAACTGATCAGAACCA 35 TCATGGGCTGGACATTGGA 16 BAX_NM_004324_human_642 CTTCTGGAGCAGGTCACAGT 16 TCTGGGACCCTGGGCCTTCTGGAGCA 36 GGTCACAGTGGTGCCCTCT 17 BAX_NM_004324_human_117 TGAGCAGATCATGAAGACAG 17 GGGGCCCACCAGCTCTGAGCAGATCA 37 TGAAGACAGGGGCCCTTTT 18 BAX_NM_004324_human_700 TATAATGCGTTTTCCTTACG 18 TCATCAGATGTGGTCTATAATGCGTTT 38 TCCTTACGTGTCTGATCA 19 BAX_NM_004324_human_673 CCCATCTTCAGATCATCAGA 19 CAGTGGTGCCCTCTCCCCATCTTCAGA 39 TCATCAGATGTGGTCTAT 20 BAX_NM_004324_human_452 AGGTGCCGGAACTGATCAGA 20 AGGCCCTGTGCACCAAGGTGCCGGAA 40 CTGATCAGAACCATCATGG Accession: NM_001188 HUGO BAK1 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 BAK1_NM_001188_human_1813 TGGTTTGTTATATCAGGGAA 41 ACAGGGCTTAGGACTTGGTTTGTTA 61 TATCAGGGAAAAGGAGTAGG  2 BAK1_NM_001188_human_911 TGGTACGAAGATTCTTCAAA 42 TGTTGGGCCAGTTTGTGGTACGAAG 62 ATTCTTCAAATCATGACTCC  3 BAK1_NM_001188_human_1820 TTATATCAGGGAAAAGGAGT 43 TTAGGACTTGGTTTGTTATATCAGG 63 GAAAAGGAGTAGGGAGTTCA  4 BAK1_NM_001188_human_1678 TCCCTTCCTCTCTCCTTATA 44 GTCCTCTCAGTTCTCTCCCTTCCTCTC 64 TCCTTATAGACACTTGCT  5 BAK1_NM_001188_human_926 TCAAATCATGACTCCCAAGG 45 TGGTACGAAGATTCTTCAAATCATG 65 ACTCCCAAGGGTGCCCTTTG  6 BAK1_NM_001188_human_1818 TGTTATATCAGGGAAAAGGA 46 GCTTAGGACTTGGTTTGTTATATCA 66 GGGAAAAGGAGTAGGGAGTT  7 BAK1_NM_001188_human_915 ACGAAGATTCTTCAAATCAT 47 GGGCCAGTTTGTGGTACGAAGATTC 67 TTCAAATCATGACTCCCAAG  8 BAK1_NM_001188_human_912 GGTACGAAGATTCTTCAAAT 48 GTTGGGCCAGTTTGTGGTACGAAGA 68 TTCTTCAAATCATGACTCCC  9 BAK1_NM_001188_human_2086 GAAGTTCTTGATTCAGCCAA 49 GGGGGTCAGGGGGGAGAAGTTCTT 69 GATTCAGCCAAATGCAGGGAG 10 BAK1_NM_001188_human_620 CCTATGAGTACTTCACCAAG 50 CCACGGCAGAGAATGCCTATGAGTA 70 CTTCACCAAGATTGCCACCA 11 BAK1_NM_001188_human_1823 TATCAGGGAAAAGGAGTAGG 51 GGACTTGGTTTGTTATATCAGGGAA 71 AAGGAGTAGGGAGTTCATCT 12 BAK1_NM_001188_human_1687 CTCTCCTTATAGACACTTGC 52 GTTCTCTCCCTTCCTCTCTCCTTATAG 72 ACACTTGCTCCCAACCCA 13 BAK1_NM_001188_human_1810 ACTTGGTTTGTTATATCAGG 53 ACTACAGGGCTTAGGACTTGGTTTG 73 TTATATCAGGGAAAAGGAGT 14 BAK1_NM_001188_human_1399 AAGATCAGCACCCTAAGAGA 54 ATTCAGCTATTCTGGAAGATCAGCA 74 CCCTAAGAGATGGGACTAGG 15 BAK1_NM_001188_human_654 GTTTGAGAGTGGCATCAATT 55 GATTGCCACCAGCCTGTTTGAGAGT 75 GGCATCAATTGGGGCCGTGT 16 BAK1_NM_001188_human_1875 GACTATCAACACCACTAGGA 56 TCTAAGTGGGAGAAGGACTATCAAC 76 ACCACTAGGAATCCCAGAGG 17 BAK1_NM_001188_human_1043 AGCTTTAGCAAGTGTGCACT 57 CCTCAAGAGTACAGAAGCTTTAGCA 77 AGTGTGCACTCCAGCTTCGG 18 BAK1_NM_001188_human_1846 TTCATCTGGAGGGTTCTAAG 58 AAAAGGAGTAGGGAGTTCATCTGG 78 AGGGTTCTAAGTGGGAGAAGG 19 BAK1_NM_001188_human_2087 AAGTTCTTGATTCAGCCAAA 59 GGGGTCAGGGGGGAGAAGTTCTTG 79 ATTCAGCCAAATGCAGGGAGG 20 BAK1_NM_001188_human_1819 GTTATATCAGGGAAAAGGAG 60 CTTAGGACTTGGTTTGTTATATCAG 80 GGAAAAGGAGTAGGGAGTTC Accession: NM_001228 HUGO CASP8 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 CASP8_NM_001228_human_2821 TTAAATCATTAGGAATTAAG 121 TCTGCTTGGATTATTTTAAATCATTAG 141 GAATTAAGTTATCTTTAA  2 CASP8_NM_001228_human_2833 GAATTAAGTTATCTTTAAAA 122 ATTTTAAATCATTAGGAATTAAGTTAT 142 CTTTAAAATTTAAGTATC  3 CASP8_NM_001228_human_2392 AACTTTAATTCTCTTTCAAA 123 TGTTAATATTCTATTAACTTTAATTCT 143 CTTTCAAAGCTAAATTCC  4 CASP8_NM_001228_human_1683 GACTGAAGTGAACTATGAAG 124 TATTCTCACCATCCTGACTGAAGTGA 144 ACTATGAAGTAAGCAACAA  5 CASP8_NM_001228_human_281 ATATTCTCCTGCCTTTTAAA 125 GGGAATATTGAGATTATATTCTCCTG 145 CCTTTTAAAAAGATGGACT  6 CASP8_NM_001228_human_2839 AGTTATCTTTAAAATTTAAG 126 AATCATTAGGAATTAAGTTATCTTTA 146 AAATTTAAGTATCTTTTTT  7 CASP8_NM_001228_human_2164 TAGATTTTCTACTTTATTAA 127 TATTTACTAATTTTCTAGATTTTCTACT 147 TTATTAATTGTTTTGCA  8 CASP8_NM_001228_human_888 CTGTGCCCAAATCAACAAGA 128 CATCCTGAAAAGAGTCTGTGCCCAAA 148 TCAACAAGAGCCTGCTGAA  9 CASP8_NM_001228_human_2283 AGCTGGTGGCAATAAATACC 129 TTTGGGAATGTTTTTAGCTGGTGGCA 149 ATAAATACCAGACACGTAC 10 CASP8_NM_001228_human_1585 TCCTACCGAAACCCTGCAGA 130 GTGAATAACTGTGTTTCCTACCGAAA 150 CCCTGCAGAGGGAACCTGG 11 CASP8_NM_001228_human_2200 TATAAGAGCTAAAGTTAAAT 131 TGTTTTGCACTTTTTTATAAGAGCTAA 151 AGTTAAATAGGATATTAA 12 CASP8_NM_001228_human_2140 CACTATGTTTATTTACTAAT 132 ACTATTTAGATATAACACTATGTTTAT 152 TTACTAATTTTCTAGATT 13 CASP8_NM_001228_human_2350 ATTGTTATCTATCAACTATA 133 GGGCTTATGATTCAGATTGTTATCTA 153 TCAACTATAAGCCCACTGT 14 CASP8_NM_001228_human_1575 TAACTGTGTTTCCTACCGAA 134 GATGGCCACTGTGAATAACTGTGTTT 154 CCTACCGAAACCCTGCAGA 15 CASP8_NM_001228_human_2397 TAATTCTCTTTCAAAGCTAA 135 ATATTCTATTAACTTTAATTCTCTTTCA 155 AAGCTAAATTCCACACT 16 CASP8_NM_001228_human_2726 TATATGCTTGGCTAACTATA 136 TGCTTTTATGATATATATATGCTTGGC 156 TAACTATATTTGCTTTTT 17 CASP8_NM_001228_human_2805 CTCTGCTTGGATTATTTTAA 137 CATTTGCTCTTTCATCTCTGCTTGGAT 157 TATTTTAAATCATTAGGA 18 CASP8_NM_001228_human_2729 ATGCTTGGCTAACTATATTT 138 TTTTATGATATATATATGCTTGGCTAA 158 CTATATTTGCTTTTTGCT 19 CASP8_NM_001228_human_2201 ATAAGAGCTAAAGTTAAATA 139 GTTTTGCACTTTTTTATAAGAGCTAAA 159 GTTAAATAGGATATTAAC 20 CASP8_NM_001228_human_2843 ATCTTTAAAATTTAAGTATC 140 ATTAGGAATTAAGTTATCTTTAAAATT 160 TAAGTATCTTTTTTCAAA Accession: NM_000675 HUGO gene ADORA2A symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 ADORA2A_NM_000675_human_2482 TAACTGCCTTTCCTTCTAAA 161 GTGAGAGGCCTTGTCTAACTGCC 181 TTTCCTTCTAAAGGGAATGTTT  2 ADORA2A_NM_000675_human_2491 TTCCTTCTAAAGGGAATGTT 162 CTTGTCTAACTGCCTTTCCTTCTAA 182 AGGGAATGTTTTTTTCTGAG  3 ADORA2A_NM_000675_human_2487 GCCTTTCCTTCTAAAGGGAA 163 AGGCCTTGTCTAACTGCCTTTCCT 183 TCTAAAGGGAATGTTTTTTTC  4 ADORA2A_NM_000675_human_2512 TTTTCTGAGATAAAATAAAA 164 CTAAAGGGAATGTTTTTTTCTGAG 184 ATAAAATAAAAACGAGCCACA  5 ADORA2A_NM_000675_human_2330 CATCTCTTGGAGTGACAAAG 165 TCTCAGTCCCAGGGCCATCTCTTG 185 GAGTGACAAAGCTGGGATCAA  6 ADORA2A_NM_000675_human_987 CATGGTGTACTTCAACTTCT 166 GGTCCCCATGAACTACATGGTGT 186 ACTTCAACTTCTTTGCCTGTGT  7 ADORA2A_NM_000675_human_2481 CTAACTGCCTTTCCTTCTAA 167 AGTGAGAGGCCTTGTCTAACTGC 187 CTTTCCTTCTAAAGGGAATGTT  8 ADORA2A_NM_000675_human_1695 CTGATGATTCATGGAGTTTG 168 TGGAGCAGGAGTGTCCTGATGAT 188 TCATGGAGTTTGCCCCTTCCTA  9 ADORA2A_NM_000675_human_264 CTCAGAGTCCTCTGTGAAAA 169 CCTGGTTTCAGGAGACTCAGAGT 189 CCTCTGTGAAAAAGCCCTTGGA 10 ADORA2A_NM_000675_human_2531 AACGAGCCACATCGTGTTTT 170 CTGAGATAAAATAAAAACGAGCC 190 ACATCGTGTTTTAAGCTTGTCC 11 ADORA2A_NM_000675_human_2492 TCCTTCTAAAGGGAATGTTT 171 TTGTCTAACTGCCTTTCCTTCTAAA 191 GGGAATGTTTTTTTCTGAGA 12 ADORA2A_NM_000675_human_978 CATGAACTACATGGTGTACT 172 TGAGGATGTGGTCCCCATGAACT 192 ACATGGTGTACTTCAACTTCTT 13 ADORA2A_NM_000675_human_2483 AACTGCCTTTCCTTCTAAAG 173 TGAGAGGCCTTGTCTAACTGCCTT 193 TCCTTCTAAAGGGAATGTTTT 14 ADORA2A_NM_000675_human_1894 CAGATGTTTCATGCTGTGAG 174 TGGGTTCTGAGGAAGCAGATGTT 194 TCATGCTGTGAGGCCTTGCACC 15 ADORA2A_NM_000675_human_976 CCCATGAACTACATGGTGTA 175 TTTGAGGATGTGGTCCCCATGAA 195 CTACATGGTGTACTTCAACTTC 16 ADORA2A_NM_000675_human_1384 AGGCAGCAAGAACCTTTCAA 176 CGCAGCCACGTCCTGAGGCAGCA 196 AGAACCTTTCAAGGCAGCTGGC 17 ADORA2A_NM_000675_human_1692 GTCCTGATGATTCATGGAGT 177 GGATGGAGCAGGAGTGTCCTGAT 197 GATTCATGGAGTTTGCCCCTTC 18 ADORA2A_NM_000675_human_993 GTACTTCAACTTCTTTGCCT 178 CATGAACTACATGGTGTACTTCAA 198 CTTCTTTGCCTGTGTGCTGGT 19 ADORA2A_NM_000675_human_2167 TGTAAGTGTGAGGAAACCCT 179 TTTTTCCAGGAAAAATGTAAGTGT 199 GAGGAAACCCTTTTTATTTTA 20 ADORA2A_NM_000675_human_1815 CCTACTTTGGACTGAGAGAA 180 TGAGGGCAGCCGGTTCCTACTTT 200 GGACTGAGAGAAGGGAGCCCCA Accession: NM_005214 HUGO CTLA4 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 CTLA4_NM_005214_human_61 TGATTCTGTGTGGGTTCAAA 201 TCTATATAAAGTCCTTGATTCTGT 221 GTGGGTTCAAACACATTTCAA  2 CTLA4_NM_005214_human_909 TTATTTGTTTGTGCATTTGG 202 GCTATCCAGCTATTTTTATTTGTTT 222 GTGCATTTGGGGGGAATTCA  3 CTLA4_NM_005214_human_1265 TGATTACATCAAGGCTTCAA 203 TCTTAAACAAATGTATGATTACAT 223 CAAGGCTTCAAAAATACTCAC  4 CTLA4_NM_005214_human_1094 GATGTGGGTCAAGGAATTAA 204 GGGATGCAGCATTATGATGTGGG 224 TCAAGGAATTAAGTTAGGGAAT  5 CTLA4_NM_005214_human_1241 CCTTTTATTTCTTAAACAAA 205 AAGTTAAATTTTATGCCTTTTATTT 225 CTTAAACAAATGTATGATTA  6 CTLA4_NM_005214_human_1266 GATTACATCAAGGCTTCAAA 206 CTTAAACAAATGTATGATTACATC 226 AAGGCTTCAAAAATACTCACA  7 CTLA4_NM_005214_human_65 TCTGTGTGGGTTCAAACACA 207 TATAAAGTCCTTGATTCTGTGTGG 227 GTTCAAACACATTTCAAAGCT  8 CTLA4_NM_005214_human_1405 TTGATAGTATTGTGCATAGA 208 TATATATATTTTAATTTGATAGTAT 228 TGTGCATAGAGCCACGTATG  9 CTLA4_NM_005214_human_1239 TGCCTTTTATTTCTTAAACA 209 TCAAGTTAAATTTTATGCCTTTTAT 229 TTCTTAAACAAATGTATGAT 10 CTLA4_NM_005214_human_1912 TCCATGAAAATGCAACAACA 210 TTTAACTCAATATTTTCCATGAAA 230 ATGCAACAACATGTATAATAT 11 CTLA4_NM_005214_human_1245 TTATTTCTTAAACAAATGTA 211 TAAATTTTATGCCTTTTATTTCTTA 231 AACAAATGTATGATTACATC 12 CTLA4_NM_005214_human_1449 TTAATGGTTTGAATATAAAC 212 GTTTTTGTGTATTTGTTAATGGTT 232 GAATATAAACACTATATGGC 13 CTLA4_NM_005214_human_1095 ATGTGGGTCAAGGAATTAAG 213 GGATGCAGCATTATGATGTGGGT 233 CAAGGAATTAAGTTAGGGAATG 14 CTLA4_NM_005214_human_1208 AGCCGAAATGATCTTTTCAA 214 GTATGAGACGTTTATAGCCGAAA 234 TGATCTTTTCAAGTTAAATTTT 15 CTLA4_NM_005214_human_1455 GTTTGAATATAAACACTATA 215 GTGTATTTGTTAATGGTTTGAATA 235 TAAACACTATATGGCAGTGTC 16 CTLA4_NM_005214_human_1237 TATGCCTTTTATTTCTTAAA 216 TTTCAAGTTAAATTTTATGCCTTTT 236 ATTTCTTAAACAAATGTATG 17 CTLA4_NM_005214_human_1911 TTCCATGAAAATGCAACAAC 217 TTTTAACTCAATATTTTCCATGAA 237 AATGCAACAACATGTATAATA 18 CTLA4_NM_005214_human_937 CATCTCTCTTTAATATAAAG 218 CATTTGGGGGGAATTCATCTCTCT 238 TTAATATAAAGTTGGATGCGG 19 CTLA4_NM_005214_human_931 GGAATTCATCTCTCTTTAAT 219 TTTGTGCATTTGGGGGGAATTCAT 239 CTCTCTTTAATATAAAGTTGG 20 CTLA4_NM_005214_human_45 ATCTATATAAAGTCCTTGAT 220 TCTGGGATCAAAGCTATCTATATA 240 AAGTCCTTGATTCTGTGTGGG Accession: NM_002286 HUGO LAG3 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 LAG3_NM_002286_human_1292 GACTTTACCCTTCGACTAGA 241 ACTGGAGACAATGGCGACTTTACC 261 CTTCGACTAGAGGATGTGAGC  2 LAG3_NM_002286_human_1096 CAACGTCTCCATCATGTATA 242 CTACAGAGATGGCTTCAACGTCTC 262 CATCATGTATAACCTCACTGT  3 LAG3_NM_002286_human_1721 GTCCTTTCTCTGCTCCTTTT 243 TTTCTCATCCTTGGTGTCCTTTCTCT 263 GCTCCTTTTGGTGACTGGA  4 LAG3_NM_002286_human_1465 TCCAGTATCTGGACAAGAAC 244 GCTTTGTGAGGTGACTCCAGTATC 264 TGGACAAGAACGCTTTGTGTG  5 LAG3_NM_002286_human_1795 ATTTTCTGCCTTAGAGCAAG 245 GTGGCGACCAAGACGATTTTCTGC 265 CTTAGAGCAAGGGATTCACCC  6 LAG3_NM_002286_human_1760 TTTCACCTTTGGAGAAGACA 246 ACTGGAGCCTTTGGCTTTCACCTTT 266 GGAGAAGACAGTGGCGACCA  7 LAG3_NM_002286_human_904 CATTTTGAACTGCTCCTTCA 247 AGCCTCCGACTGGGTCATTTTGAA 267 CTGCTCCTTCAGCCGCCCTGA   8 LAG3_NM_002286_human_1398 TCATCACAGTGACTCCCAAA 248 CTGTCACATTGGCAATCATCACAGT 268 GACTCCCAAATCCTTTGGGT  9 LAG3_NM_002286_human_1758 GCTTTCACCTTTGGAGAAGA 249 TGACTGGAGCCTTTGGCTTTCACCT 269 TTGGAGAAGACAGTGGCGAC 10 LAG3_NM_002286_human_1753 CTTTGGCTTTCACCTTTGGA 250 TTTGGTGACTGGAGCCTTTGGCTTT 270 CACCTTTGGAGAAGACAGTG 11 LAG3_NM_002286_human_905 ATTTTGAACTGCTCCTTCAG 251 GCCTCCGACTGGGTCATTTTGAACT 271 GCTCCTTCAGCCGCCCTGAC 12 LAG3_NM_002286_human_1387 CACATTGGCAATCATCACAG 252 GCTCAATGCCACTGTCACATTGGC 272 AATCATCACAGTGACTCCCAA 13 LAG3_NM_002286_human_301 TTTCTGACCTCCTTTTGGAG 253 ACTGCCCCCTTTCCTTTTCTGACCTC 273 CTTTTGGAGGGCTCAGCGC 14 LAG3_NM_002286_human_895 CGACTGGGTCATTTTGAACT 254 ATCTCTCAGAGCCTCCGACTGGGT 274 CATTTTGAACTGCTCCTTCAG 15 LAG3_NM_002286_human_1625 TACTTCACAGAGCTGTCTAG 255 CTTGGAGCAGCAGTGTACTTCACA 275 GAGCTGTCTAGCCCAGGTGCC 16 LAG3_NM_002286_human_1390 ATTGGCAATCATCACAGTGA 256 CAATGCCACTGTCACATTGGCAATC 276 ATCACAGTGACTCCCAAATC 17 LAG3_NM_002286_human_1703 CTGTTTCTCATCCTTGGTGT 257 GCAGGCCACCTCCTGCTGTTTCTCA 277 TCCTTGGTGTCCTTTCTCTG 18 LAG3_NM_002286_human_1453 TTGTGAGGTGACTCCAGTAT 258 CCTGGGGAAGCTGCTTTGTGAGGT 278 GACTCCAGTATCTGGACAAGA 19 LAG3_NM_002286_human_1754 TTTGGCTTTCACCTTTGGAG 259 TTGGTGACTGGAGCCTTTGGCTTTC 279 ACCTTTGGAGAAGACAGTGG 20 LAG3_NM_002286_human_1279 TGGAGACAATGGCGACTTTA 260 TGACCTCCTGGTGACTGGAGACAA 280 TGGCGACTTTACCCTTCGACT Accession: NM_005018 HUGO PDCD1 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 PDCDLN1_NM_005018_human_2070 TATTATATTATAATTATAAT 281 CCTTCCCTGTGGTTCTATTATATTAT 301 AATTATAATTAAATATGAG  2 PDCDLN1_NM_005018_human_2068 TCTATTATATTATAATTATA 282 CCCCTTCCCTGTGGTTCTATTATATT 302 ATAATTATAATTAAATATG  3 PDCDLN1_NM_005018_human_1854 CATTCCTGAAATTATTTAAA 283 GCTCTCCTTGGAACCCATTCCTGAA 303 ATTATTTAAAGGGGTTGGCC  4 PDCDLN1_NM_005018_human_2069 CTATTATATTATAATTATAA 284 CCCTTCCCTGTGGTTCTATTATATT 304 ATAATTATAATTAAATATGA  5 PDCDLN1_NM_005018_human_1491 AGTTTCAGGGAAGGTCAGAA 285 CTGCAGGCCTAGAGAAGTTTCAGG 305 GAAGGTCAGAAGAGCTCCTGG  6 PDCDLN1_NM_005018_human_2062 TGTGGTTCTATTATATTATA 286 GGGATCCCCCTTCCCTGTGGTTCTA 306 TTATATTATAATTATAATTA  7 PDCDLN1_NM_005018_human_719 TGTGTTCTCTGTGGACTATG 287 CCCCTCAGCCGTGCCTGTGTTCTCT 307 GTGGACTATGGGGAGCTGGA  8 PDCDLN1_NM_005018_human_1852 CCCATTCCTGAAATTATTTA 288 GAGCTCTCCTTGGAACCCATTCCTG 308 AAATTATTTAAAGGGGTTGG  9 PDCDLN1_NM_005018_human_1490 TGCCACCATTGTCTTTCCTA 289 TGAGCAGACGGAGTATGCCACCAT 309 TGTCTTTCCTAGCGGAATGGG 10 PDCDLN1_NM_005018_human_812 AAGTTTCAGGGAAGGTCAGA 290 CCTGCAGGCCTAGAGAAGTTTCAG 310 GGAAGGTCAGAAGAGCTCCTG 11 PDCDLN1_NM_005018_human_2061 CTGTGGTTCTATTATATTAT 291 AGGGATCCCCCTTCCCTGTGGTTCT 311 ATTATATTATAATTATAATT 12 PDCDLN1_NM_005018_human_2067 TTCTATTATATTATAATTAT 292 CCCCCTTCCCTGTGGTTCTATTATA 312 TTATAATTATAATTAAATAT 13 PDCDLN1_NM_005018_human_1493 TTTCAGGGAAGGTCAGAAGA 293 GCAGGCCTAGAGAAGTTTCAGGG 313 AAGGTCAGAAGAGCTCCTGGCT 14 PDCDLN1_NM_005018_human_1845 CTTGGAACCCATTCCTGAAA 294 ACCCTGGGAGCTCTCCTTGGAACC 314 CATTCCTGAAATTATTTAAAG 15 PDCDLN1_NM_005018_human_2058 TCCCTGTGGTTCTATTATAT 295 ACAAGGGATCCCCCTTCCCTGTGG 315 TTCTATTATATTATAATTATA 16 PDCDLN1_NM_005018_human_2060 CCTGTGGTTCTATTATATTA 296 AAGGGATCCCCCTTCCCTGTGGTTC 316 TATTATATTATAATTATAAT 17 PDCDLN1_NM_005018_human_1847 TGGAACCCATTCCTGAAATT 297 CCTGGGAGCTCTCCTTGGAACCCA 317 TTCCTGAAATTATTTAAAGGG 18 PDCDLN1_NM_005018_human_2055 CCTTCCCTGTGGTTCTATTA 298 GGGACAAGGGATCCCCCTTCCCTG 318 TGGTTCTATTATATTATAATT 19 PDCDLN1_NM_005018_human_2057 TTCCCTGTGGTTCTATTATA 299 GACAAGGGATCCCCCTTCCCTGTG 319 GTTCTATTATATTATAATTAT 20 PDCDLN1_NM_005018_human_1105 CACAGGACTCATGTCTCAAT 300 CAGGCACAGCCCCACCACAGGACT 320 CATGTCTCAATGCCCACAGTG Accession: NM_004612 HUGO TGFBR1 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 TGFBRL1_NM_004612_human_5263 CCTGTTTATTACAACTTAAA 321 GTTAATAACATTCAACCTGTTTAT 341 TACAACTTAAAAGGAACTTCA  2 TGFBRL1_NM_004612_human_1323 CCATTGGTGGAATTCATGAA 322 TTGCTCGACGATGTTCCATTGGTG 342 GAATTCATGAAGATTACCAAC  3 TGFBRL1_NM_004612_human_6389 TTTTCCTTATAACAAAGACA 323 TTTAGGGATTTTTTTTTTTCCTTAT 343 AACAAAGACATCACCAGGAT  4 TGFBRL1_NM_004612_human_3611 TGTATTACTTGTTTAATAAT 324 TTTTTATAGTTGTGTTGTATTACTT 344 GTTTAATAATAATCTCTAAT  5 TGFBRL1_NM_004612_human_3882 TTATTGAATCAAAGATTGAG 325 TGCTGAAGATATTTTTTATTGAAT 345 CAAAGATTGAGTTACAATTAT  6 TGFBRL1_NM_004612_human_3916 TTCTTACCTAAGTGGATAAA 326 GTTACAATTATACTTTTCTTACCTA 346 AGTGGATAAAATGTACTTTT  7 TGFBRL1_NM_004612_human_5559 ATGTTGCTCAGTTACTCAAA 327 TAAAGTATGGGTATTATGTTGCTC 347 AGTTACTCAAATGGTACTGTA  8 TGFBRL1_NM_004612_human_5595 ATATTTGTACCCCAAATAAC 328 GGTACTGTATTGTTTATATTTGTA 348 CCCCAAATAACATCGTCTGTA  9 TGFBRL1_NM_004612_human_5222 TGTAAATGTAAACTTCTAAA 329 TTATGCAATCTTGTTTGTAAATGT 349 AAACTTCTAAAAATATGGTTA 10 TGFBRL1_NM_004612_human_3435 AGAATGAGTGACATATTACA 330 AACCAAAGTAATTTTAGAATGAG 350 TGACATATTACATAGGAATTTA 11 TGFBRL1_NM_004612_human_3709 CCATTTCTAAGCCTACCAGA 331 GTTGTTGTTTTTGGGCCATTTCTA 351 AGCCTACCAGATCTGCTTTAT 12 TGFBRL1_NM_004612_human_5826 ATATTCCAAAAGAATGTAAA 332 ATTGTATTTGTAGTAATATTCCAA 352 AAGAATGTAAATAGGAAATAG 13 TGFBRL1_NM_004612_human_3146 TTACTTCCAATGCTATGAAG 333 TATAATAACTGGTTTTTACTTCCA 353 ATGCTATGAAGTCTCTGCAGG 14 TGFBRL1_NM_004612_human_2675 TCTTTATCTGTTCAAAGACT 334 TGTAAGCCATTTTTTTCTTTATCTG 354 TTCAAAGACTTATTTTTTAA 15 TGFBRL1_NM_004612_human_2529 GTCTAAGTATACTTTTAAAA 335 CATTTTAATTGTGTTGTCTAAGTA 355 TACTTTTAAAAAATCAAGTGG 16 TGFBRL1_NM_004612_human_5079 ATCTTTGGACATGTACTGCA 336 GAGATACTAAGGATTATCTTTGG 356 ACATGTACTGCAGCTTCTTGTC 17 TGFBRL1_NM_004612_human_3607 GTGTTGTATTACTTGTTTAA 337 TTTGTTTTTATAGTTGTGTTGTATT 357 ACTTGTTTAATAATAATCTC 18 TGFBRL1_NM_004612_human_5994 TGCTGTAGATGGCAACTAGA 338 CATGCCATATGTAGTTGCTGTAGA 358 TGGCAACTAGAACCTTTGAGT 19 TGFBRL1_NM_004612_human_2177 TCTTTCACTTATTCAGAACA 339 GTATACTATTATTGTTCTTTCACTT 359 ATTCAGAACATTACATGCCT 20 TGFBRL1_NM_004612_human_5814 GTATTTGTAGTAATATTCCA 340 TTTAAATTGTATATTGTATTTGTA 360 GTAATATTCCAAAAGAATGTA Accession: NM_032782 HUGO HAVCR2 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 HAVCR2_NM_032782_human_937 CTCATAGCAAAGAGAAGATA 361 TTTTCAAATGGTATTCTCATAGCA 381 AAGAGAAGATACAGAATTTAA  2 HAVCR2_NM_032782_human_932 GTATTCTCATAGCAAAGAGA 362 TTTAATTTTCAAATGGTATTCTCAT 382 AGCAAAGAGAAGATACAGAA  3 HAVCR2_NM_032782_human_2126 TTGCTTGTTGTGTGCTTGAA 363 TGTATTGGCCAAGTTTTGCTTGTT 383 GTGTGCTTGAAAGAAAATATC  4 HAVCR2_NM_032782_human_2171 TATTCGTGGACCAAACTGAA 364 TCTGACCAACTTCTGTATTCGTGG 384 ACCAAACTGAAGCTATATTTT  5 HAVCR2_NM_032782_human_158 ATTGTGGAGTAGACAGTTGG 365 GCTACTGCTCATGTGATTGTGGA 385 GTAGACAGTTGGAAGAAGTACC  6 HAVCR2_NM_032782_human_2132 GTTGTGTGCTTGAAAGAAAA 366 GGCCAAGTTTTGCTTGTTGTGTGC 386 TTGAAAGAAAATATCTCTGAC  7 HAVCR2_NM_032782_human_2131 TGTTGTGTGCTTGAAAGAAA 367 TGGCCAAGTTTTGCTTGTTGTGTG 387 CTTGAAAGAAAATATCTCTGA  8 HAVCR2_NM_032782_human_2313 CCCTAAACTTAAATTTCAAG 368 TTGACAGAGAGTGGTCCCTAAAC 388 TTAAATTTCAAGACGGTATAGG  9 HAVCR2_NM_032782_human_489 ACATCCAGATACTGGCTAAA 369 GATGTGAATTATTGGACATCCAG 389 ATACTGGCTAAATGGGGATTTC 10 HAVCR2_NM_032782_human_1272 CATTTTCAGAAGATAATGAC 370 GGAGCAGAGTTTTCCCATTTTCAG 390 AAGATAATGACTCACATGGGA 11 HAVCR2_NM_032782_human_785 CACATTGGCCAATGAGTTAC 371 TCTAACACAAATATCCACATTGGC  391 CAATGAGTTACGGGACTCTAG 12 HAVCR2_NM_032782_human_2127 TGCTTGTTGTGTGCTTGAAA 372 GTATTGGCCAAGTTTTGCTTGTTG 392 TGTGCTTGAAAGAAAATATCT 13 HAVCR2_NM_032782_human_164 GAGTAGACAGTTGGAAGAAG 373 GCTCATGTGATTGTGGAGTAGAC 393 AGTTGGAAGAAGTACCCAGTCC 14 HAVCR2_NM_032782_human_2130 TTGTTGTGTGCTTGAAAGAA 374 TTGGCCAAGTTTTGCTTGTTGTGT 394 GCTTGAAAGAAAATATCTCTG 15 HAVCR2_NM_032782_human_911 CGGCGCTTTAATTTTCAAAT 375 TCTGGCTCTTATCTTCGGCGCTTT 395 AATTTTCAAATGGTATTCTCA 16 HAVCR2_NM_032782_human_1543 TTTGGCACAGAAAGTCTAAA 376 TGAAAGCATAACTTTTTTGGCACA 396 GAAAGTCTAAAGGGGCCACTG 17 HAVCR2_NM_032782_human_2346 GATCTGTCTTGCTTATTGTT 377 AGACGGTATAGGCTTGATCTGTC 397 TTGCTTATTGTTGCCCCCTGCG 18 HAVCR2_NM_032782_human_2107 GGTGTGTATTGGCCAAGTTT 378 GAAGTGCATTTGATTGGTGTGTA 398 TTGGCCAAGTTTTGCTTGTTGT 19 HAVCR2_NM_032782_human_1270 CCCATTTTCAGAAGATAATG 379 ATGGAGCAGAGTTTTCCCATTTTC 399 AGAAGATAATGACTCACATGG 20 HAVCR2_NM_032782_human_1545 TGGCACAGAAAGTCTAAAGG 380 AAAGCATAACTTTTTTGGCACAGA 400 AAGTCTAAAGGGGCCACTGAT Accession: NM_002987 HUGO CCL17 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 CCL17_NM_002987_human_385 AAATACCTGCAAAGCCTTGA 401 GTGAAGAATGCAGTTAAATACCTGC 421 AAAGCCTTGAGAGGTCTTGA  2 CCL17_NM_002987_human_318 TTTTGTAACTGTGCAGGGCA 402 CAGGGATGCCATCGTTTTTGTAACT 422 GTGCAGGGCAGGGCCATCTG  3 CCL17_NM_002987_human_367 AGAGTGAAGAATGCAGTTAA 403 GACCCCAACAACAAGAGAGTGAAG 423 AATGCAGTTAAATACCTGCAA  4 CCL17_NM_002987_human_396 AAGCCTTGAGAGGTCTTGAA 404 AGTTAAATACCTGCAAAGCCTTGAG 424 AGGTCTTGAAGCCTCCTCAC  5 CCL17_NM_002987_human_386 AATACCTGCAAAGCCTTGAG 405 TGAAGAATGCAGTTAAATACCTGCA 425 AAGCCTTGAGAGGTCTTGAA  6 CCL17_NM_002987_human_378 TGCAGTTAAATACCTGCAAA 406 CAAGAGAGTGAAGAATGCAGTTAA 426 ATACCTGCAAAGCCTTGAGAG  7 CCL17_NM_002987_human_357 CAACAACAAGAGAGTGAAGA 407 CATCTGTTCGGACCCCAACAACAAG 427 AGAGTGAAGAATGCAGTTAA  8 CCL17_NM_002987_human_55 CTGAATTCAAAACCAGGGTG 408 CTGCTGATGGGAGAGCTGAATTCAA 428 AACCAGGGTGTCTCCCTGAG  9 CCL17_NM_002987_human_387 ATACCTGCAAAGCCTTGAGA 409 GAAGAATGCAGTTAAATACCTGCAA 429 AGCCTTGAGAGGTCTTGAAG 10 CCL17_NM_002987_human_254 TTCCCCTTAGAAAGCTGAAG 410 ACTTCAAGGGAGCCATTCCCCTTAG 430 AAAGCTGAAGACGTGGTACC 11 CCL17_NM_002987_human_49 GGAGAGCTGAATTCAAAACC 411 CACCGCCTGCTGATGGGAGAGCTG 431 AATTCAAAACCAGGGTGTCTC 12 CCL17_NM_002987_human_379 GCAGTTAAATACCTGCAAAG 412 AAGAGAGTGAAGAATGCAGTTAAA 432 TACCTGCAAAGCCTTGAGAGG 13 CCL17_NM_002987_human_372 GAAGAATGCAGTTAAATACC 413 CAACAACAAGAGAGTGAAGAATGC 433 AGTTAAATACCTGCAAAGCCT 14 CCL17_NM_002987_human_377 ATGCAGTTAAATACCTGCAA 414 ACAAGAGAGTGAAGAATGCAGTTA 434 AATACCTGCAAAGCCTTGAGA 15 CCL17_NM_002987_human_252 CATTCCCCTTAGAAAGCTGA 415 GTACTTCAAGGGAGCCATTCCCCTT 435 AGAAAGCTGAAGACGTGGTA 16 CCL17_NM_002987_human_51 AGAGCTGAATTCAAAACCAG 416 CCGCCTGCTGATGGGAGAGCTGAAT 436 TCAAAACCAGGGTGTCTCCC 17 CCL17_NM_002987_human_45 GATGGGAGAGCTGAATTCAA 417 GTGTCACCGCCTGCTGATGGGAGA 437 GCTGAATTCAAAACCAGGGTG 18 CCL17_NM_002987_human_44 TGATGGGAGAGCTGAATTCA 418 AGTGTCACCGCCTGCTGATGGGAGA 438 GCTGAATTCAAAACCAGGGT 19 CCL17_NM_002987_human_16 ACTTTGAGCTCACAGTGTCA 419 GCTCAGAGAGAAGTGACTTTGAGCT 439 CACAGTGTCACCGCCTGCTG 20 CCL17_NM_002987_human_368 GAGTGAAGAATGCAGTTAAA 420 ACCCCAACAACAAGAGAGTGAAGA 440 ATGCAGTTAAATACCTGCAAA Accession: NM_002990 HUGO CCL22 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 CCL22_NM_002990_human_2083 GTATTTGAAAACAGAGTAAA 441 GCTGGAGTTATATATGTATTTGAA 461 AACAGAGTAAATACTTAAGAG  2 CCL22_NM_002990_human_298 CAATAAGCTGAGCCAATGAA 442 GGTGAAGATGATTCTCAATAAGC 462 TGAGCCAATGAAGAGCCTACTC    3 CCL22_NM_002990_human_2103 TACTTAAGAGGCCAAATAGA 443 TGAAAACAGAGTAAATACTTAAG 463 AGGCCAAATAGATGAATGGAAG  4 CCL22_NM_002990_human_2081 ATGTATTTGAAAACAGAGTA 444 AAGCTGGAGTTATATATGTATTTG 464 AAAACAGAGTAAATACTTAAG  5 CCL22_NM_002990_human_2496 TTCATACAGCAAGTATGGGA 445 TTGAGAAATATTCTTTTCATACAG 465 CAAGTATGGGACAGCAGTGTC  6 CCL22_NM_002990_human_1052 CTGCAGACAAAATCAATAAA 446 GAGCCCAGAAAGTGGCTGCAGAC 466 AAAATCAATAAAACTAATGTCC  7 CCL22_NM_002990_human_1053 TGCAGACAAAATCAATAAAA 447 AGCCCAGAAAGTGGCTGCAGACA 467 AAATCAATAAAACTAATGTCCC  8 CCL22_NM_002990_human_2112 GGCCAAATAGATGAATGGAA 448 AGTAAATACTTAAGAGGCCAAAT 468 AGATGAATGGAAGAATTTTAGG  9 CCL22_NM_002990_human_299 AATAAGCTGAGCCAATGAAG 449 GTGAAGATGATTCTCAATAAGCT 469 GAGCCAATGAAGAGCCTACTCT 10 CCL22_NM_002990_human_2108 AAGAGGCCAAATAGATGAAT 450 ACAGAGTAAATACTTAAGAGGCC 470 AAATAGATGAATGGAAGAATTT 11 CCL22_NM_002990_human_2116 AAATAGATGAATGGAAGAAT 451 AATACTTAAGAGGCCAAATAGAT 471 GAATGGAAGAATTTTAGGAACT 12 CCL22_NM_002990_human_2091 AAACAGAGTAAATACTTAAG 452 TATATATGTATTTGAAAACAGAGT 472 AAATACTTAAGAGGCCAAATA 13 CCL22_NM_002990_human_2067 AGCTGGAGTTATATATGTAT 453 TGACTTGGTATTATAAGCTGGAG 473 TTATATATGTATTTGAAAACAG 14 CCL22_NM_002990_human_2047 ACCTTTGACTTGGTATTATA 454 ATGGTGTGAAAGACTACCTTTGA 474 CTTGGTATTATAAGCTGGAGTT 15 CCL22_NM_002990_human_238 AACCTTCAGGGATAAGGAGA 455 TGGCGTGGTGTTGCTAACCTTCA 475 GGGATAAGGAGATCTGTGCCGA 16 CCL22_NM_002990_human_2037 GTGAAAGACTACCTTTGACT 456 AATTCATGCTATGGTGTGAAAGA 476 CTACCTTTGACTTGGTATTATA 17 CCL22_NM_002990_human_2030 CTATGGTGTGAAAGACTACC 457 ACAATCAAATTCATGCTATGGTGT 477 GAAAGACTACCTTTGACTTGG 18 CCL22_NM_002990_human_1682 CACTACGGCTGGCTAATTTT 458 ATTACAGGTGTGTGCCACTACGG 478 CTGGCTAATTTTTGTATTTTTA 19 CCL22_NM_002990_human_2071 GGAGTTATATATGTATTTGA 459 TTGGTATTATAAGCTGGAGTTATA 479 TATGTATTTGAAAACAGAGTA 20 CCL22_NM_002990_human_1111 ATATCAATACAGAGACTCAA 460 CCAAAAGGCAGTTACATATCAAT 480 ACAGAGACTCAAGGTCACTAGA Accession: NM_005618 HUGO DLL1 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 DLL1_NM_005618_human_3246 CTGTTTTGTTAATGAAGAAA 481 TATTTGAGTTTTTTACTGTTTTGTTA 501 ATGAAGAAATTCCTTTTTA  2 DLL1_NM_005618_human_3193 TTGTATATAAATGTATTTAT 482 TGTGACTATATTTTTTTGTATATAAA 502 TGTATTTATGGAATATTGT  3 DLL1_NM_005618_human_3247 TGTTTTGTTAATGAAGAAAT 483 ATTTGAGTTTTTTACTGTTTTGTTAA 503 TGAAGAAATTCCTTTTTAA  4 DLL1_NM_005618_human_3141 AATTTTGGTAAATATGTACA 484 GTTTTTTATAATTTAAATTTTGGTAA 504 ATATGTACAAAGGCACTTC  5 DLL1_NM_005618_human_3293 AAATTTTATGAATGACAAAA 485 ATATTTTTCCAAAATAAATTTTATGA 505 ATGACAAAAAAAAAAAAAA  6 DLL1_NM_005618_human_3208 TTTATGGAATATTGTGCAAA 486 TTGTATATAAATGTATTTATGGAATA 506 TTGTGCAAATGTTATTTGA  7 DLL1_NM_005618_human_3243 TTACTGTTTTGTTAATGAAG 487 TGTTATTTGAGTTTTTTACTGTTTTGT 507 TAATGAAGAAATTCCTTT  8 DLL1_NM_005618_human_2977 TTCTTGAATTAGAAACACAA 488 TTATGAGCCAGTCTTTTCTTGAATTA 508 GAAACACAAACACTGCCTT  9 DLL1_NM_005618_human_2874 CAGTTGCTCTTAAGAGAATA 489 CCGTTGCACTATGGACAGTTGCTCTT 509 AAGAGAATATATATTTAAA 10 DLL1_NM_005618_human_2560 CAACTTCAAAAGACACCAAG 490 CGGACTCGGGCTGTTCAACTTCAAA 510 AGACACCAAGTACCAGTCGG 11 DLL1_NM_005618_human_3285 TCCAAAATAAATTTTATGAA 491 TTTTTAAAATATTTTTCCAAAATAAA 511 TTTTATGAATGACAAAAAA 12 DLL1_NM_005618_human_2909 GAACTGAATTACGCATAAGA 492 TATATTTAAATGGGTGAACTGAATT 512 ACGCATAAGAAGCATGCACT 13 DLL1_NM_005618_human_1173 GGATTTTGTGACAAACCAGG 493 TGTGATGAGCAGCATGGATTTTGTG 513 ACAAACCAGGGGAATGCAAG 14 DLL1_NM_005618_human_3244 TACTGTTTTGTTAATGAAGA 494 GTTATTTGAGTTTTTTACTGTTTTGTT 514 AATGAAGAAATTCCTTTT 15 DLL1_NM_005618_human_3144 TTTGGTAAATATGTACAAAG 495 TTTTATAATTTAAATTTTGGTAAATA 515 TGTACAAAGGCACTTCGGG 16 DLL1_NM_005618_human_3286 CCAAAATAAATTTTATGAAT 496 TTTTAAAATATTTTTCCAAAATAAAT 516 TTTATGAATGACAAAAAAA 17 DLL1_NM_005618_human_3133 ATAATTTAAATTTTGGTAAA 497 TGATGTTCGTTTTTTATAATTTAAAT 517 TTTGGTAAATATGTACAAA 18 DLL1_NM_005618_human_2901 AAATGGGTGAACTGAATTAC 498 AGAGAATATATATTTAAATGGGTGA 518 ACTGAATTACGCATAAGAAG 19 DLL1_NM_005618_human_3168 TTCGGGTCTATGTGACTATA 499 TATGTACAAAGGCACTTCGGGTCTA 519 TGTGACTATATTTTTTTGTA 20 DLL1_NM_005618_human_3245 ACTGTTTTGTTAATGAAGAA 500 TTATTTGAGTTTTTTACTGTTTTGTTA 520 ATGAAGAAATTCCTTTTT Accession: NM_000639 HUGO FASLG gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 FASLG_NM_000639_human_1154 TAGCTCCTCAACTCACCTAA 521 GGTTCAAAATGTCTGTAGCTCCTC 541 AACTCACCTAATGTTTATGAG  2 FASLG_NM_000639_human_1771 ATGTTTTCCTATAATATAAT 522 TGTCAGCTACTAATGATGTTTTCC 542 TATAATATAATAAATATTTAT  3 FASLG_NM_000639_human_1774 TTTTCCTATAATATAATAAA 523 CAGCTACTAATGATGTTTTCCTAT 543 AATATAATAAATATTTATGTA  4 FASLG_NM_000639_human_1776 TTCCTATAATATAATAAATA 524 GCTACTAATGATGTTTTCCTATAA 544 TATAATAAATATTTATGTAGA  5 FASLG_NM_000639_human_1086 TGCATTTGAGGTCAAGTAAG 525 GAGGGTCTTCTTACATGCATTTGA 545 GGTCAAGTAAGAAGACATGAA  6 FASLG_NM_000639_human_1750 ATTGATTGTCAGCTACTAAT 526 TAGTGCTTAAAAATCATTGATTGT 546 CAGCTACTAATGATGTTTTCC  7 FASLG_NM_000639_human_1820 AAATGAAAACATGTAATAAA 527 ATGTGCATTTTTGTGAAATGAAAA 547 CATGTAATAAAAAGTATATGT  8 FASLG_NM_000639_human_1659 ATTGTGAAGTACATATTAGG 528 AGAGAGAATGTAGATATTGTGAA 548 GTACATATTAGGAAAATATGGG  9 FASLG_NM_000639_human_667 GCTTTCTGGAGTGAAGTATA 529 CTATGGAATTGTCCTGCTTTCTGG 549 AGTGAAGTATAAGAAGGGTGG 10 FASLG_NM_000639_human_1692 CATTTGGTCAAGATTTTGAA 530 GGAAAATATGGGTTGCATTTGGT 550 CAAGATTTTGAATGCTTCCTGA 11 FASLG_NM_000639_human_986 GGCTTATATAAGCTCTAAGA 531 TCTCAGACGTTTTTCGGCTTATAT 551 AAGCTCTAAGAGAAGCACTTT 12 FASLG_NM_000639_human_911 ACCAGTGCTGATCATTTATA 532 GCAGTGTTCAATCTTACCAGTGCT  552 GATCATTTATATGTCAACGTA 13 FASLG_NM_000639_human_598 CCATTTAACAGGCAAGTCCA 533 GCTGAGGAAAGTGGCCCATTTAA 553 CAGGCAAGTCCAACTCAAGGTC 14 FASLG_NM_000639_human_1665 AAGTACATATTAGGAAAATA 534 AATGTAGATATTGTGAAGTACAT 554 ATTAGGAAAATATGGGTTGCAT 15 FASLG_NM_000639_human_1625 TGTGTGTGTGTATGACTAAA 535 GTGTGTGTGTGTGTGTGTGTGTG 555 TGTATGACTAAAGAGAGAATGT 16 FASLG_NM_000639_human_1238 AAGAGGGAGAAGCATGAAAA 536 CTGGGCTGCCATGTGAAGAGGGA 556 GAAGCATGAAAAAGCAGCTACC 17 FASLG_NM_000639_human_1632 GTGTATGACTAAAGAGAGAA 537 TGTGTGTGTGTGTGTGTGTATGA 557 CTAAAGAGAGAATGTAGATATT 18 FASLG_NM_000639_human_1581 GTATTTCCAGTGCAATTGTA 538 CCTAACACAGCATGTGTATTTCCA 558 GTGCAATTGTAGGGGTGTGTG 19 FASLG_NM_000639_human_1726 CAACTCTAATAGTGCTTAAA 539 ATGCTTCCTGACAATCAACTCTAA 559 TAGTGCTTAAAAATCATTGAT 20 FASLG_NM_000639_human_1626 GTGTGTGTGTATGACTAAAG 540 TGTGTGTGTGTGTGTGTGTGTGT 560 GTATGACTAAAGAGAGAATGTA Accession: NM_001267706 HUGO CD274 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 CD274_NM_001267706_human_3222 ACCTGCATTAATTTAATAAA 561 ATTGTCACTTTTTGTACCTGCATTA 581 ATTTAATAAAATATTCTTAT  2 CD274_NM_001267706_human_1538 AACTTGCCCAAACCAGTAAA 562 GCAAACAGATTAAGTAACTTGCC 582 CAAACCAGTAAATAGCAGACCT  3 CD274_NM_001267706_human_1218 ATTTGCTCACATCTAGTAAA 563 ACTTGCTGCTTAATGATTTGCTCA 583 CATCTAGTAAAACATGGAGTA  4 CD274_NM_001267706_human_1998 CCTTTGCCATATAATCTAAT 564 TTTATTCCTGATTTGCCTTTGCCAT 584 ATAATCTAATGCTTGTTTAT  5 CD274_NM_001267706_human_2346 ATATAGCAGATGGAATGAAT 565 ATTTTAGTGTTTCTTATATAGCAG 585 ATGGAATGAATTTGAAGTTCC  6 CD274_NM_001267706_human_1997 GCCTTTGCCATATAATCTAA 566 ATTTATTCCTGATTTGCCTTTGCCA 586 TATAATCTAATGCTTGTTTA  7 CD274_NM_001267706_human_1992 GATTTGCCTTTGCCATATAA 567 ATTATATTTATTCCTGATTTGCCTT 587 TGCCATATAATCTAATGCTT  8 CD274_NM_001267706_human_1905 AATTTTCATTTACAAAGAGA 568 CTTAATAATCAGAGTAATTTTCAT 588 TTACAAAGAGAGGTCGGTACT  9 CD274_NM_001267706_human_2336 AGTGTTTCTTATATAGCAGA 569 ATTTTTATTTATTTTAGTGTTTCTT 589 ATATAGCAGATGGAATGAAT 10 CD274_NM_001267706_human_2656 GCTTTCTGTCAAGTATAAAC 570 GAACTTTTGTTTTCTGCTTTCTGTC 590 AAGTATAAACTTCACTTTGA 11 CD274_NM_001267706_human_2235 CATTTGGAAATGTATGTTAA 571 TCTAAAGATAGTCTACATTTGGAA 591 ATGTATGTTAAAAGCACGTAT 12 CD274_NM_001267706_human_2329 TTATTTTAGTGTTTCTTATA 572 CTTTGCTATTTTTATTTATTTTAGT 592 GTTTCTTATATAGCAGATGG 13 CD274_NM_001267706_human_1433 GTGGTAGCCTACACACATAA 573 CAGCTTTACAATTATGTGGTAGCC 593 TACACACATAATCTCATTTCA 14 CD274_NM_001267706_human_1745 ATGAGGAGATTAACAAGAAA 574 GGAGCTCATAGTATAATGAGGAG 594 ATTAACAAGAAAATGTATTATT 15 CD274_NM_001267706_human_1183 CAATTTTGTCGCCAAACTAA 575 TTGTAGTAGATGTTACAATTTTGT 595 CGCCAAACTAAACTTGCTGCT 16 CD274_NM_001267706_human_2345 TATATAGCAGATGGAATGAA 576 TATTTTAGTGTTTCTTATATAGCA  596 GATGGAATGAATTTGAAGTTC 17 CD274_NM_001267706_human_2069 AAATGCCACTAAATTTTAAA 577 CTGTCTTTTCTATTTAAATGCCACT 597 AAATTTTAAATTCATACCTT 18 CD274_NM_001267706_human_2414 TCTTTCCCATAGCTTTTCAT 578 TTTGTTTCTAAGTTATCTTTCCCAT 598 AGCTTTTCATTATCTTTCAT 19 CD274_NM_001267706_human_129 TATATTCATGACCTACTGGC 579 GATATTTGCTGTCTTTATATTCAT 599 GACCTACTGGCATTTGCTGAA 20 CD274_NM_001267706_human_1783 GTCCAGTGTCATAGCATAAG 580 TATTATTACAATTTAGTCCAGTGT 600 CATAGCATAAGGATGATGCGA Accession: NM_002164 HUGO gene IDO1 symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 IDO1_NM_002164_human_1896 ATTCTGTCATAATAAATAAA 601 AAAAAAAAAAGATATATTCTGTCA 621 TAATAAATAAAAATGCATAAG  2 IDO1_NM_002164_human_1532 TATCTTATCATTGGAATAAA 602 AAGTTTTGTAATCTGTATCTTATCA 622 TTGGAATAAAATGACATTCA  3 IDO1_NM_002164_human_578 GTGATGGAGACTGCAGTAAA 603 TTTTGTTCTCATTTCGTGATGGAGA 623 CTGCAGTAAAGGATTCTTCC  4 IDO1_NM_002164_human_1897 TTCTGTCATAATAAATAAAA 604 AAAAAAAAAGATATATTCTGTCAT 624 AATAAATAAAAATGCATAAGA  5 IDO1_NM_002164_human_1473 CTTGTAGGAAAACAACAAAA 605 AATACCTGTGCATTTCTTGTAGGAA 625 AACAACAAAAGGTAATTATG  6 IDO1_NM_002164_human_1547 ATAAAATGACATTCAATAAA 606 TATCTTATCATTGGAATAAAATGAC 626 ATTCAATAAATAAAAATGCA  7 IDO1_NM_002164_human_412 CGTAAGGTCTTGCCAAGAAA 607 GGTCATGGAGATGTCCGTAAGGTC 627 TTGCCAAGAAATATTGCTGTT  8 IDO1_NM_002164_human_1472 TCTTGTAGGAAAACAACAAA 608 AAATACCTGTGCATTTCTTGTAGGA 628 AAACAACAAAAGGTAATTAT  9 IDO1_NM_002164_human_1248 AACTGGAGGCACTGATTTAA 609 ACTGGAAGCCAAAGGAACTGGAG 629 GCACTGATTTAATGAATTTCCT 10 IDO1_NM_002164_human_1440 CAATACAAAAGACCTCAAAA 610 GTTTTACCAATAATGCAATACAAAA 630 GACCTCAAAATACCTGTGCA 11 IDO1_NM_002164_human_636 TGCTTCTGCAATCAAAGTAA 611 GGTGGAAATAGCAGCTGCTTCTGC 631 AATCAAAGTAATTCCTACTGT 12 IDO1_NM_002164_human_1551 AATGACATTCAATAAATAAA 612 TTATCATTGGAATAAAATGACATTC 632 AATAAATAAAAATGCATAAG 13 IDO1_NM_002164_human_1538 ATCATTGGAATAAAATGACA 613 TGTAATCTGTATCTTATCATTGGAA 633 TAAAATGACATTCAATAAAT 14 IDO1_NM_002164_human_1430 ACCAATAATGCAATACAAAA 614 ACTATGCAATGTTTTACCAATAATG 634 CAATACAAAAGACCTCAAAA 15 IDO1_NM_002164_human_1527 ATCTGTATCTTATCATTGGA 615 ACTAGAAGTTTTGTAATCTGTATCT 635 TATCATTGGAATAAAATGAC 16 IDO1_NM_002164_human_1533 ATCTTATCATTGGAATAAAA 616 AGTTTTGTAATCTGTATCTTATCAT 636 TGGAATAAAATGACATTCAA 17 IDO1_NM_002164_human_632 CAGCTGCTTCTGCAATCAAA 617 TATTGGTGGAAATAGCAGCTGCTT 637 CTGCAATCAAAGTAATTCCTA 18 IDO1_NM_002164_human_1439 GCAATACAAAAGACCTCAAA 618 TGTTTTACCAATAATGCAATACAAA 638 AGACCTCAAAATACCTGTGC 19 IDO1_NM_002164_human_657 TCCTACTGTATTCAAGGCAA 619 TGCAATCAAAGTAATTCCTACTGTA 639 TTCAAGGCAATGCAAATGCA 20 IDO1_NM_002164_human_1398 CAGAGCCACAAACTAATACT 620 CATTACCCATTGTAACAGAGCCAC AAACTAATACTATGCAATGTT Accession: NM_001558 HUGO gene IL10RA symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 IL10RA_NM_001558_human_3364 TTGTTCATTTATTTATTGGA 641 CTTTATTTATTTATTTTGTTCATTT 661 ATTTATTGGAGAGGCAGCAT  2 IL10RA_NM_001558_human_3626 TTATTCCAATAAATTGTCAA 642 AGTGATACATGTTTTTTATTCCAA 662 TAAATTGTCAAGACCACAGGA  3 IL10RA_NM_001558_human_2395 TATTTTCTGGACACTCAAAC 643 AGATCTTAAGGTATATATTTTCTG 663 GACACTCAAACACATCATAAT  4 IL10RA_NM_001558_human_3375 TTTATTGGAGAGGCAGCATT 644 TATTTTGTTCATTTATTTATTGGAG 664 AGGCAGCATTGCACAGTGAA  5 IL10RA_NM_001558_human_3469 ACCTTGGAGAAGTCACTTAT 645 GTTTCCAGTGGTATGACCTTGGA 665 GAAGTCACTTATCCTCTTGGAG  6 IL10RA_NM_001558_human_3351 TTATTTATTTATTTTGTTCA 646 GTTCCCTTGAAAGCTTTATTTATTT 666 ATTTTGTTCATTTATTTATT  7 IL10RA_NM_001558_human_2108 CTCTTTCCTGTATCATAAAG 647 TCTCCCTCCTAGGAACTCTTTCCT 667 GTATCATAAAGGATTATTTGC  8 IL10RA_NM_001558_human_3563 CTGAGGAAATGGGTATGAAT 648 GGATGTGAGGTTCTGCTGAGGAA 668 ATGGGTATGAATGTGCCTTGAA  9 IL10RA_NM_001558_human_3579 GAATGTGCCTTGAACACAAA 649 TGAGGAAATGGGTATGAATGTGC 669 CTTGAACACAAAGCTCTGTCAA 10 IL10RA_NM_001558_human_2403 GGACACTCAAACACATCATA 650 AGGTATATATTTTCTGGACACTCA 670 AACACATCATAATGGATTCAC 11 IL10RA_NM_001558_human_2115 CTGTATCATAAAGGATTATT 651 CCTAGGAACTCTTTCCTGTATCAT 671 AAAGGATTATTTGCTCAGGGG 12 IL10RA_NM_001558_human_563 TCACTTCCGAGAGTATGAGA 652 TGAAAGCATCTTCAGTCACTTCCG 672 AGAGTATGAGATTGCCATTCG 13 IL10RA_NM_001558_human_3197 TCTCTGGAGCATTCTGAAAA 653 TCTCAGCCCTGCCTTTCTCTGGAG 673 CATTCTGAAAACAGATATTCT 14 IL10RA_NM_001558_human_2987 TTATGCCAGAGGCTAACAGA 654 AAGCTGGCTTGTTTCTTATGCCAG 674 AGGCTAACAGATCCAATGGGA 15 IL10RA_NM_001558_human_1278 AGTGGCATTGACTTAGTTCA 655 AGGGGCCAGGATGACAGTGGCA 675 TTGACTTAGTTCAAAACTCTGAG 16 IL10RA_NM_001558_human_2398 TTTCTGGACACTCAAACACA 656 TCTTAAGGTATATATTTTCTGGAC 676 ACTCAAACACATCATAATGGA 17 IL10RA_NM_001558_human_3390 GCATTGCACAGTGAAAGAAT 657 TTTATTGGAGAGGCAGCATTGCA 677 CAGTGAAAGAATTCTGGATATC 18 IL10RA_NM_001558_human_3468 GACCTTGGAGAAGTCACTTA 658 TGTTTCCAGTGGTATGACCTTGGA 678 GAAGTCACTTATCCTCTTGGA 19 IL10RA_NM_001558_human_610 TCACGTTCACACACAAGAAA 659 AGGTGCCGGGAAACTTCACGTTC 679 ACACACAAGAAAGTAAAACATG 20 IL10RA_NM_001558_human_3446 ACTTTGCTGTTTCCAGTGGT 660 GAAATTCTAGCTCTGACTTTGCTG 680 TTTCCAGTGGTATGACCTTGG Accession: NM_000214 HUGO gene JAG1 symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 JAG1_NM_000214_human_4799 TATTTGATTTATTAACTTAA 681 ATTAATCACTGTGTATATTTGATTT 701 ATTAACTTAATAATCAAGAG  2 JAG1_NM_000214_human_5658 GAAAAGTAATATTTATTAAA 682 TTGGCAATAAATTTTGAAAAGTAA 702 TATTTATTAAATTTTTTTGTA  3 JAG1_NM_000214_human_4752 ACTTTGTATAGTTATGTAAA 683 AATGTCAAAAGTAGAACTTTGTAT 703 AGTTATGTAAATAATTCTTTT  4 JAG1_NM_000214_human_5418 GAATACTTGAACCATAAAAT 684 TCTAATAAGCTAGTTGAATACTTGA 704 ACCATAAAATGTCCAGTAAG  5 JAG1_NM_000214_human_5641 TCTTGGCAATAAATTTTGAA 685 TCTTTGATGTGTTGTTCTTGGCAAT 705 AAATTTTGAAAAGTAATATT  6 JAG1_NM_000214_human_5150 TTTCTGCTTTAGACTTGAAA 686 TGTTTGTTTTTTGTTTTTCTGCTTTA 706 GACTTGAAAAGAGACAGGC  7 JAG1_NM_000214_human_4526 TATATTTATTGACTCTTGAG 687 GATCATAGTTTTATTTATATTTATT 707 GACTCTTGAGTTGTTTTTGT  8 JAG1_NM_000214_human_4566 TATGATGACGTACAAGTAGT 688 TTTGTATATTGGTTTTATGATGACG 708 TACAAGTAGTTCTGTATTTG  9 JAG1_NM_000214_human_5634 GTGTTGTTCTTGGCAATAAA 689 AAATGCATCTTTGATGTGTTGTTCT 709 TGGCAATAAATTTTGAAAAG 10 JAG1_NM_000214_human_173 CTGATCTAAAAGGGAATAAA 690 CCTTTTTCCATGCAGCTGATCTAAA 710 AGGGAATAAAAGGCTGCGCA 11 JAG1_NM_000214_human_5031 TACGACGTCAGATGTTTAAA 691 GATGGAATTTTTTTGTACGACGTCA 711 GATGTTTAAAACACCTTCTA 12 JAG1_NM_000214_human_4817 AATAATCAAGAGCCTTAAAA 692 TTGATTTATTAACTTAATAATCAAG 712 AGCCTTAAAACATCATTCCT 13 JAG1_NM_000214_human_5685 GTATGAAAACATGGAACAGT 693 TTATTAAATTTTTTTGTATGAAAAC 713 ATGGAACAGTGTGGCCTCTT 14 JAG1_NM_000214_human_4560 TGGTTTTATGATGACGTACA 694 GTTGTTTTTGTATATTGGTTTTATG 714 ATGACGTACAAGTAGTTCTG 15 JAG1_NM_000214_human_5151 TTCTGCTTTAGACTTGAAAA 695 GTTTGTTTTTTGTTTTTCTGCTTTAG 715 ACTTGAAAAGAGACAGGCA 16 JAG1_NM_000214_human_5642 CTTGGCAATAAATTTTGAAA 696 CTTTGATGTGTTGTTCTTGGCAATA 716 AATTTTGAAAAGTAATATTT 17 JAG1_NM_000214_human_5377 TTTAATCTACTGCATTTAGG 697 GATTTGATTTTTTTTTTTAATCTACT 717 GCATTTAGGGAGTATTCTA 18 JAG1_NM_000214_human_4756 TGTATAGTTATGTAAATAAT 698 TCAAAAGTAGAACTTTGTATAGTTA 718 TGTAAATAATTCTTTTTTAT 19 JAG1_NM_000214_human_4523 ATTTATATTTATTGACTCTT 699 TTAGATCATAGTTTTATTTATATTTA 719 TTGACTCTTGAGTTGTTTT 20 JAG1_NM_000214_human_5325 CTTTTCACCATTCGTACATA 700 TGTAAATTCTGATTTCTTTTCACCAT 720 TCGTACATAATACTGAACC Accession: NM_002226 HUGO JAG2 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 JAG2_NM_002226_human_4266 CGTTTCTTTAACCTTGTATA 721 AATGTTTATTTTCTACGTTTCTTTAA 741 CCTTGTATAAATTATTCAG  2 JAG2_NM_002226_human_5800 TAAATGAATGAACGAATAAA 722 GGCAGAACAAATGAATAAATGAAT 742 GAACGAATAAAAATTTTGACC  3 JAG2_NM_002226_human_5450 TCATTCATTTATTCCTTTGT 723 GGTCAAAATTTTTATTCATTCATTT 743 ATTCCTTTGTTTTGCTTGGT   4 JAG2_NM_002226_human_5021 GTAAATGTGTACATATTAAA 724 TGAAAGTGCATTTTTGTAAATGTGT 744 ACATATTAAAGGAAGCACTC  5 JAG2_NM_002226_human_5398 ACCCACGAATACGTATCAAG 725 AGTATAAAATTGCTTACCCACGAAT 745 ACGTATCAAGGTCTTAAGGA  6 JAG2_NM_002226_human_5371 GTTTTATAAAATAGTATAAA 726 AAACAGCTGAAAACAGTTTTATAA 746 AATAGTATAAAATTGCTTACC  7 JAG2_NM_002226_human_5691 CAACTGAGTCAAGGAGCAAA 727 TGAGGGGTAGGAGGTCAACTGAG 747 TCAAGGAGCAAAGCCAAGAACC  8 JAG2_NM_002226_human_5025 ATGTGTACATATTAAAGGAA 728 AGTGCATTTTTGTAAATGTGTACAT 748 ATTAAAGGAAGCACTCTGTA  9 JAG2_NM_002226_human_4269 TTCTTTAACCTTGTATAAAT 729 GTTTATTTTCTACGTTTCTTTAACCT 749 TGTATAAATTATTCAGTAA 10 JAG2_NM_002226_human_4258 ATTTTCTACGTTTCTTTAAC 730 AAAAACCAAATGTTTATTTTCTACG 750 TTTCTTTAACCTTGTATAAA 11 JAG2_NM_002226_human_5369 CAGTTTTATAAAATAGTATA 731 TAAAACAGCTGAAAACAGTTTTAT 751 AAAATAGTATAAAATTGCTTA 12 JAG2_NM_002226_human_5780 GCACAGGCAGAACAAATGAA 732 GAGTGAGGCTGCCTTGCACAGGCA 752 GAACAAATGAATAAATGAATG 13 JAG2_NM_002226_human_4302 TCAGGCTGAAAACAATGGAG 733 ATTATTCAGTAACTGTCAGGCTGA 753 AAACAATGGAGTATTCTCGGA 14 JAG2_NM_002226_human_5387 TAAAATTGCTTACCCACGAA 734 TTTTATAAAATAGTATAAAATTGCT 754 TACCCACGAATACGTATCAA 15 JAG2_NM_002226_human_4301 GTCAGGCTGAAAACAATGGA 735 AATTATTCAGTAACTGTCAGGCTG 755 AAAACAATGGAGTATTCTCGG 16 JAG2_NM_002226_human_5023 AAATGTGTACATATTAAAGG 736 AAAGTGCATTTTTGTAAATGTGTAC 756 ATATTAAAGGAAGCACTCTG 17 JAG2_NM_002226_human_4293 CAGTAACTGTCAGGCTGAAA 737 CTTGTATAAATTATTCAGTAACTGT 757 CAGGCTGAAAACAATGGAGT 18 JAG2_NM_002226_human_4321 GTATTCTCGGATAGTTGCTA 738 GCTGAAAACAATGGAGTATTCTCG 758 GATAGTTGCTATTTTTGTAAA 19 JAG2_NM_002226_human_3994 TCTCACACAAATTCACCAAA 739 AGGCGGAGAAGTTCCTCTCACACA 759 AATTCACCAAAGATCCTGGCC 20 JAG2_NM_002226_human_5466 TTGTTTTGCTTGGTCATTCA 740 CATTCATTTATTCCTTTGTTTTGCTT 760 GGTCATTCAGAGGCAAGGT Accession: NM_001315 HUGO gene MAPK14 symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 MAPK14_NM_001315_human_670 TCATGCGAAAAGAACCTACA 761 ATTTCAGTCCATCATTCATGCGAAAA 781 GAACCTACAGAGAACTGCG  2 MAPK14_NM_001315_human_833 AAATGTCAGAAGCTTACAGA 762 CTGAACAACATTGTGAAATGTCAGA 782 AGCTTACAGATGACCATGTT  3 MAPK14_NM_001315_human_707 AAACATATGAAACATGAAAA 763 GAACTGCGGTTACTTAAACATATGA 783 AACATGAAAATGTGATTGGT  4 MAPK14_NM_001315_human_863 CAGTTCCTTATCTACCAAAT 764 ACAGATGACCATGTTCAGTTCCTTAT 784 CTACCAAATTCTCCGAGGT  5 MAPK14_NM_001315_human_1150 TCCTGGTACAGACCATATTA 765 TGGAAGAACATTGTTTCCTGGTACA 785 GACCATATTAACCAGCTTCA  6 MAPK14_NM_001315_human_866 TTCCTTATCTACCAAATTCT 766 GATGACCATGTTCAGTTCCTTATCTA 786 CCAAATTCTCCGAGGTCTA  7 MAPK14_NM_001315_human_1149 TTCCTGGTACAGACCATATT 767 CTGGAAGAACATTGTTTCCTGGTAC 787 AGACCATATTAACCAGCTTC  8 MAPK14_NM_001315_human_896 AAGTATATACATTCAGCTGA 768 ATTCTCCGAGGTCTAAAGTATATAC 788 ATTCAGCTGACATAATTCAC  9 MAPK14_NM_001315_human_1076 CATTACAACCAGACAGTTGA 769 ATGCTGAACTGGATGCATTACAACC 789 AGACAGTTGATATTTGGTCA 10 MAPK14_NM_001315_human_926 AGGGACCTAAAACCTAGTAA 770 GCTGACATAATTCACAGGGACCTAA 790 AACCTAGTAATCTAGCTGTG 11 MAPK14_NM_001315_human_765 CTCTGGAGGAATTCAATGAT 771 TTACACCTGCAAGGTCTCTGGAGGA 791 ATTCAATGATGTGTATCTGG 12 MAPK14_NM_001315_human_706 TAAACATATGAAACATGAAA 772 AGAACTGCGGTTACTTAAACATATG 792 AAACATGAAAATGTGATTGG 13 MAPK14_NM_001315_human_815 GATCTGAACAACATTGTGAA 773 CATCTCATGGGGGCAGATCTGAACA 793 ACATTGTGAAATGTCAGAAG 14 MAPK14_NM_001315_human_862 TCAGTTCCTTATCTACCAAA 774 TACAGATGACCATGTTCAGTTCCTTA 794 TCTACCAAATTCTCCGAGG 15 MAPK14_NM_001315_human_917 ATAATTCACAGGGACCTAAA 775 ATACATTCAGCTGACATAATTCACA 795 GGGACCTAAAACCTAGTAAT 16 MAPK14_NM_001315_human_887 CGAGGTCTAAAGTATATACA 776 ATCTACCAAATTCTCCGAGGTCTAA 796 AGTATATACATTCAGCTGAC 17 MAPK14_NM_001315_human_832 GAAATGTCAGAAGCTTACAG 777 TCTGAACAACATTGTGAAATGTCAG 797 AAGCTTACAGATGACCATGT 18 MAPK14_NM_001315_human_1125 AGCTGTTGACTGGAAGAACA 778 GATGCATAATGGCCGAGCTGTTGAC 798 TGGAAGAACATTGTTTCCTG 19 MAPK14_NM_001315_human_879 AAATTCTCCGAGGTCTAAAG 779 AGTTCCTTATCTACCAAATTCTCCGA 799 GGTCTAAAGTATATACATT 20 MAPK14_NM_001315_human_725 AATGTGATTGGTCTGTTGGA 780 CATATGAAACATGAAAATGTGATTG 800 GTCTGTTGGACGTTTTTACA Accession: NM_003745 HUGO SOCS1 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 SOCS1_NM_003745_human_1141 CTGCTGTGCAGAATCCTATT 801 TCTGGCTTTATTTTTCTGCTGTGCA  821 GAATCCTATTTTATATTTTT  2 SOCS1_NM_003745_human_1143 GCTGTGCAGAATCCTATTTT 802 TGGCTTTATTTTTCTGCTGTGCAGA 822 ATCCTATTTTATATTTTTTA  3 SOCS1_NM_003745_human_1170 TTAAAGTCAGTTTAGGTAAT 803 CCTATTTTATATTTTTTAAAGTCAG 823 TTTAGGTAATAAACTTTATT  4 SOCS1_NM_003745_human_1144 CTGTGCAGAATCCTATTTTA 804 GGCTTTATTTTTCTGCTGTGCAGAA 824 TCCTATTTTATATTTTTTAA  5 SOCS1_NM_003745_human_1076 GTTTACATATACCCAGTATC 805 CTCCTACCTCTTCATGTTTACATAT 825 ACCCAGTATCTTTGCACAAA  6 SOCS1_NM_003745_human_837 ATTTTGTTATTACTTGCCTG 806 CTGGGATGCCGTGTTATTTTGTTA 826 TTACTTGCCTGGAACCATGTG  7 SOCS1_NM_003745_human_819 TAACTGGGATGCCGTGTTAT 807 CCGTGCACGCAGCATTAACTGGGATG 827 CCGTGTTATTTTGTTATTA  8 SOCS1_NM_003745_human_841 TGTTATTACTTGCCTGGAAC 808 GATGCCGTGTTATTTTGTTATTACT 828 TGCCTGGAACCATGTGGGTA  9 SOCS1_NM_003745_human_1138 TTTCTGCTGTGCAGAATCCT 809 GTCTCTGGCTTTATTTTTCTGCTGT 829 GCAGAATCCTATTTTATATT 10 SOCS1_NM_003745_human_831 CGTGTTATTTTGTTATTACT 810 CATTAACTGGGATGCCGTGTTATTTTG 830 TTATTACTTGCCTGGAAC 11 SOCS1_NM_003745_human_1168 TTTTAAAGTCAGTTTAGGTA 811 ATCCTATTTTATATTTTTTAAAGTC 831 AGTTTAGGTAATAAACTTTA 12 SOCS1_NM_003745_human_1142 TGCTGTGCAGAATCCTATTT 812 CTGGCTTTATTTTTCTGCTGTGCAG 832 AATCCTATTTTATATTTTTT 13 SOCS1_NM_003745_human_825 GGATGCCGTGTTATTTTGTT 813 ACGCAGCATTAACTGGGATGCCGTGTT 833 ATTTTGTTATTACTTGCC 14 SOCS1_NM_003745_human_1169 TTTAAAGTCAGTTTAGGTAA 814 TCCTATTTTATATTTTTTAAAGTCA 834 GTTTAGGTAATAAACTTTAT 15 SOCS1_NM_003745_human_1171 TAAAGTCAGTTTAGGTAATA 815 CTATTTTATATTTTTTAAAGTCAGT 835 TTAGGTAATAAACTTTATTA 16 SOCS1_NM_003745_human_1140 TCTGCTGTGCAGAATCCTAT 816 CTCTGGCTTTATTTTTCTGCTGTGC 836 AGAATCCTATTTTATATTTT 17 SOCS1_NM_003745_human_1082 ATATACCCAGTATCTTTGCA 817 CCTCTTCATGTTTACATATACCCA 837 GTATCTTTGCACAAACCAGGG 18 SOCS1_NM_003745_human_1150 AGAATCCTATTTTATATTTT 818 ATTTTTCTGCTGTGCAGAATCCTAT 838 TTTATATTTTTTAAAGTCAG 19 SOCS1_NM_003745_human_1011 GGTTGTTGTAGCAGCTTAAC 819 CCTCTGGGTCCCCCTGGTTGTTGTAGC 839 AGCTTAACTGTATCTGGA 20 SOCS1_NM_003745_human_1087 CCCAGTATCTTTGCACAAAC 820 TCATGTTTACATATACCCAGTAT 840 CTTTGCACAAACCAGGGGTTGG Accession: NM_003150 HUGO STAT3 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 STAT3_NM_003150_human_4897 ATATTGCTGTATCTACTTTA 841 TTTTTTTTTTTTGGTATATTGCTGT 861 ATCTACTTTAACTTCCAGAA  2 STAT3_NM_003150_human_4325 TGTTTGTTAAATCAAATTAG 842 GTTTCTGTGGAATTCTGTTTGTTA 862 AATCAAATTAGCTGGTCTCTG  3 STAT3_NM_003150_human_2730 TTTATCTAAATGCAAATAAG 843 TGTGGGTGATCTGCTTTTATCTAA 863 ATGCAAATAAGGATGTGTTCT  4 STAT3_NM_003150_human_3615 ATTTTCCTTTGTAATGTATT 844 TTTATAAATAGACTTATTTTCCTTT 864 GTAATGTATTGGCCTTTTAG  5 STAT3_NM_003150_human_453 TATCAGCACAATCTACGAAG 845 GAGTCGAATGTTCTCTATCAGCAC 865 AATCTACGAAGAATCAAGCAG  6 STAT3_NM_003150_human_4477 AGCTTAACTGATAAACAGAA 846 CTTCAGTACATAATAAGCTTAACT 866 GATAAACAGAATATTTAGAAA  7 STAT3_NM_003150_human_2870 GTTGTTGTTGTTCTTAGACA 847 CAGCTTTTTGTTATTGTTGTTGTTG 867 TTCTTAGACAAGTGCCTCCT  8 STAT3_NM_003150_human_2873 GTTGTTGTTCTTAGACAAGT 848 CTTTTTGTTATTGTTGTTGTTGTTC 868 TTAGACAAGTGCCTCCTGGT  9 STAT3_NM_003150_human_3096 TCTGTATTTAAGAAACTTAA 849 TATCAGCATAGCCTTTCTGTATTT 869 AAGAAACTTAAGCAGCCGGGC 10 STAT3_NM_003150_human_3613 TTATTTTCCTTTGTAATGTA 850 TTTTTATAAATAGACTTATTTTCCT 870 TTGTAATGTATTGGCCTTTT 11 STAT3_NM_003150_human_4481 TAACTGATAAACAGAATATT 851 AGTACATAATAAGCTTAACTGATA 871 AACAGAATATTTAGAAAGGTG 12 STAT3_NM_003150_human_1372 ACATTCTGGGCACAAACACA 852 GATCCCGGAAATTTAACATTCTGG 872 GCACAAACACAAAAGTGATGA 13 STAT3_NM_003150_human_2720 GTGATCTGCTTTTATCTAAA 853 AATGAGTGAATGTGGGTGATCTG 873 CTTTTATCTAAATGCAAATAAG 14 STAT3_NM_003150_human_1044 CAGACCCGTCAACAAATTAA 854 GCAGAATCTCAACTTCAGACCCGT 874 CAACAAATTAAGAAACTGGAG 15 STAT3_NM_003150_human_1148 GGAGCTGTTTAGAAACTTAA 855 GGAGGAGAGAATCGTGGAGCTG 875 TTTAGAAACTTAATGAAAAGTGC 16 STAT3_NM_003150_human_4523 ACCATTGGGTTTAAATCATA 856 GTGAGACTTGGGCTTACCATTGG 876 GTTTAAATCATAGGGACCTAGG 17 STAT3_NM_003150_human_3573 GGAGAATCTAAGCATTTTAG 857 AATAGGAAGGTTTAAGGAGAATC 877 TAAGCATTTTAGACTTTTTTTT 18 STAT3_NM_003150_human_2987 CCTTGCTGACATCCAAATAG 858 CATTGCACTTTTTAACCTTGCTGA 878 CATCCAAATAGAAGATAGGAC 19 STAT3_NM_003150_human_3041 AAATTAAGAAATAATAACAA 859 CCTAGGTTTCTTTTTAAATTAAGA 879 AATAATAACAATTAAAGGGCA 20 STAT3_NM_003150_human_3037 TTTTAAATTAAGAAATAATA 860 AAGCCCTAGGTTTCTTTTTAAATT 880 AAGAAATAATAACAATTAAAG Accession: NM_006290 HUGO TNFAIP3 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 TNFAIP3_NM_006290_human_3451 AGCTTGAACTGAGGAGTAAA 881 ACTTCTAAAGAAGTTAGCTTGAAC 901 TGAGGAGTAAAAGTGTGTACA  2 TNFAIP3_NM_006290_human_916 CCTTTGCAACATCCTCAGAA 882 AATACACATATTTGTCCTTTGCAA 902 CATCCTCAGAAGGCCAATCAT  3 TNFAIP3_NM_006290_human_4422 TTCTTTCCAAAGATACCAAA 883 ACGAATCTTTATAATTTCTTTCCAA 903 AGATACCAAATAAACTTCAG  4 TNFAIP3_NM_006290_human_3688 TTATTTTATTACAAACTTCA 884 TGTAATTCACTTTATTTATTTTATT 904 ACAAACTTCAAGATTATTTA  5 TNFAIP3_NM_006290_human_4536 TATTTATACTTATTATAAAA 885 GTGAAAAAAAGTAATTATTTATAC 905 TTATTATAAAAAGTATTTGAA  6 TNFAIP3_NM_006290_human_949 CATTTCAGACAAAATGCTAA 886 AAGGCCAATCATTGTCATTTCAGA 906 CAAAATGCTAAGAAGTTTGGA  7 TNFAIP3_NM_006290_human_1214 ATGAAGGAGAAGCTCTTAAA 887 GATCCTGAAAATGAGATGAAGGA 907 GAAGCTCTTAAAAGAGTACTTA  8 TNFAIP3_NM_006290_human_4489 ATTTTGTGTTGATCATTATT 888 AGTTGATATCTTAATATTTTGTGT 908 TGATCATTATTTCCATTCTTA  9 TNFAIP3_NM_006290_human_2204 TTCATCGAGTACAGAGAAAA 889 TTTTGCACACTGTGTTTCATCGAG 909 TACAGAGAAAACAAACATTTT 10 TNFAIP3_NM_006290_human_3394 TTACTGGGAAGACGTGTAAC 890 AAAAATTAGAATATTTTACTGGGA 910 AGACGTGTAACTCTTTGGGTT 11 TNFAIP3_NM_006290_human_2355 TCATTGAAGCTCAGAATCAG 891 ACTGCCAGAAGTGTTTCATTGAA 911 GCTCAGAATCAGAGATTTCATG 12 TNFAIP3_NM_006290_human_4508 TTCCATTCTTAATGTGAAAA 892 TGTGTTGATCATTATTTCCATTCTT 912 AATGTGAAAAAAAGTAATTA 13 TNFAIP3_NM_006290_human_2332 TGAAGGATACTGCCAGAAGT 893 TGGAAGCACCATGTTTGAAGGAT 913 ACTGCCAGAAGTGTTTCATTGA 14 TNFAIP3_NM_006290_human_4650 CACAAGAGTCAACATTAAAA 894 ATAAATGTAACTTTTCACAAGAGT 914 CAACATTAAAAAATAAATTAT 15 TNFAIP3_NM_006290_human_4533 AATTATTTATACTTATTATA 895 AATGTGAAAAAAAGTAATTATTTA 915 TACTTATTATAAAAAGTATTT 16 TNFAIP3_NM_006290_human_3907 TTCGTGCTTCTCCTTATGAA 896 CATATTCATCGATGTTTCGTGCTT 916 CTCCTTATGAAACTCCAGCTA 17 TNFAIP3_NM_006290_human_3689 TATTTTATTACAAACTTCAA 897 GTAATTCACTTTATTTATTTTATTA 917 CAAACTTCAAGATTATTTAA 18 TNFAIP3_NM_006290_human_3694 TATTACAAACTTCAAGATTA 898 TCACTTTATTTATTTTATTACAAAC 918 TTCAAGATTATTTAAGTGAA 19 TNFAIP3_NM_006290_human_4467 CTCTTAAAGTTGATATCTTA 899 TGTTTTCATCTAATTCTCTTAAAGT 919 TGATATCTTAATATTTTGTG 20 TNFAIP3_NM_006290_human_4426 TTCCAAAGATACCAAATAAA 900 ATCTTTATAATTTCTTTCCAAAGAT 920 ACCAAATAAACTTCAGTGTT Accession: NM_003326 HUGO gene TNFSF4 symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 TNFSF4_NM_003326_human_2984 AATTTGACTTAGCCACTAAC 921 GAGATCAGAATTTTAAATTTGACT 941 TAGCCACTAACTAGCCATGTA  2 TNFSF4_NM_003326__human_3422 GATATTAATAATATAGTTAA 922 GAGAGTATTAATATTGATATTAAT 942 AATATAGTTAATAGTAATATT  3 TNFSF4_NM_003326_human_3119 CTGTGAATGCACATATTAAA 923 TGCTTACAGTGTTATCTGTGAATG 943 CACATATTAAATGTCTATGTT  4 TNFSF4_NM_003326_human_2208 GTTTTCTATTTCCTCTTAAG 924 GGATTTTTTTTTCCTGTTTTCTATT 944 TCCTCTTAAGTACACCTTCA  5 TNFSF4_NM_003326_human_1727 AAATAGCACTAAGAAGTTAT 925 ATTCAATCTGATGTCAAATAGCAC  945 TAAGAAGTTATTGTGCCTTAT  6 TNFSF4_NM_003326_human_3311 CCAATCCCGATCCAAATCAT 926 AATGCTTAAGGGATTCCAATCCC 946 GATCCAAATCATAATTTGTTCT  7 TNFSF4_NM_003326_human_3286 CTATTTAGAGAATGCTTAAG 927 TTAGTTAGATATTTTCTATTTAGA 947 GAATGCTTAAGGGATTCCAAT  8 TNFSF4_NM_003326_human_1222 CAGTTTGCATATTGCCTAAA 928 AGGTTAAATTGATTGCAGTTTGCA 948 TATTGCCTAAATTTAAACTTT  9 TNFSF4_NM_003326_human_326 CTCGAATTCAAAGTATCAAA 929 TATCACATCGGTATCCTCGAATTC 949 AAAGTATCAAAGTACAATTTA 10 TNFSF4_NM_003326_human_3117 ATCTGTGAATGCACATATTA 930 TATGCTTACAGTGTTATCTGTGAA 950 TGCACATATTAAATGTCTATG 11 TNFSF4_NM_003326_human_2938 TTTGTGGGAAAAGAATTGAA 931 TATACATGGCAGAGTTTTGTGGG 951 AAAAGAATTGAATGAAAAGTCA 12 TNFSF4_NM_003326_human_2537 ATTGACCATGTTCTGCAAAA 932 ATTTCACTTTTTGTTATTGACCATG 952 TTCTGCAAAATTGCAGTTAC 13 TNFSF4_NM_003326_human_776 GATTCTTCATTGCAAGTGAA 933 GGTGGACAGGGCATGGATTCTTC 953 ATTGCAAGTGAAGGAGCCTCCC 14 TNFSF4_NM_003326_human_1721 GATGTCAAATAGCACTAAGA 934 TATCAAATTCAATCTGATGTCAAA 954 TAGCACTAAGAAGTTATTGTG 15 TNFSF4_NM_003326_human_1459 GTATACAGGGAGAGTGAGAT 935 AAGAGAGATTTTCTTGTATACAG 955 GGAGAGTGAGATAACTTATTGT 16 TNFSF4_NM_003326_human_3152 GTTGCTATGAGTCAAGGAGT 936 AATGTCTATGTTCTTGTTGCTATG 956 AGTCAAGGAGTGTAACCTTCT 17 TNFSF4_NM_003326_human_1882 TAGTTGAAATGTCCCCTTAA 937 GTATCCCCTTATGTTTAGTTGAAA 957 TGTCCCCTTAACTTGATATAA 18 TNFSF4_NM_003326_human_1980 CTCTGTGCCAAACCTTTTAT 938 GATGATTTGTAACTTCTCTGTGCC  958 AAACCTTTTATAAACATAAAT 19 TNFSF4_NM_003326_human_1770 CTCTGTCTAGAAATACCATA 939 ATGAAAAATAATGATCTCTGTCTA 959 GAAATACCATAGACCATATAT 20 TNFSF4_NM_003326_human_1680 GGTTTCAAGAAATGAGGTGA 940 CACAGAAACATTGCTGGTTTCAA 960 GAAATGAGGTGATCCTATTATC Accession: NM_006293 HUGO TYRO3 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 TYRO3_NM_006293_human_3927 AGTTGCTGTTTAAAATAGAA 961 CATTTCCAAGCTGTTAGTTGCTGTT  981 TAAAATAGAAATAAAATTGA  2 TYRO3_NM_006293_human_3932 CTGTTTAAAATAGAAATAAA 962 CCAAGCTGTTAGTTGCTGTTTAAAA  982 TAGAAATAAAATTGAAGACT  3 TYRO3_NM_006293_human_1731 GGCATCAGCGATGAACTAAA 963 ACATTGGACAGCTTGGGCATCAGC  983 GATGAACTAAAGGAAAAACTG  4 TYRO3_NM_006293_human_3699 AATATCCTAAGACTAACAAA 964 GCTACCAAATCTCAAAATATCCTAA  984 GACTAACAAAGGCAGCTGTG  5 TYRO3_NM_006293_human_3928 GTTGCTGTTTAAAATAGAAA 965 ATTTCCAAGCTGTTAGTTGCTGTTT  985 AAAATAGAAATAAAATTGAA  6 TYRO3_NM_006293_human_3938 AAAATAGAAATAAAATTGAA 966 TGTTAGTTGCTGTTTAAAATAGAAA  986 TAAAATTGAAGACTAAAGAC  7 TYRO3_NM_006293_human_842 CTGTGAAGCTCACAACCTAA 967 GAGCACCATGTTTTCCTGTGAAGC  987 TCACAACCTAAAAGGCCTGGC  8 TYRO3_NM_006293_human_3953 TTGAAGACTAAAGACCTAAA 968 AAAATAGAAATAAAATTGAAGACT  988 AAAGACCTAAAAAAAAAAAAA  9 TYRO3_NM_006293_human_3703 TCCTAAGACTAACAAAGGCA 969 CCAAATCTCAAAATATCCTAAGACT  989 AACAAAGGCAGCTGTGTCTG 10 TYRO3_NM_006293_human_3909 GGACATTTCCAAGCTGTTAG 970 GGTCCTAGCTGTTAGGGACATTTC  990 CAAGCTGTTAGTTGCTGTTTA 11 TYRO3_NM_006293_human_3190 ATGTTTCCATGGTTACCATG 971 AGGAGTGGGGTGGTTATGTTTCCA  991 TGGTTACCATGGGTGTGGATG 12 TYRO3_NM_006293_human_3926 TAGTTGCTGTTTAAAATAGA 972 ACATTTCCAAGCTGTTAGTTGCTGT  992 TTAAAATAGAAATAAAATTG 13 TYRO3_NM_006293_human_3949 AAAATTGAAGACTAAAGACC 973 GTTTAAAATAGAAATAAAATTGAA  993 GACTAAAGACCTAAAAAAAAA 14 TYRO3_NM_006293_human_3900 AGCTGTTAGGGACATTTCCA 974 CATGGGGCGGGTCCTAGCTGTTAG  994 GGACATTTCCAAGCTGTTAGT 15 TYRO3_NM_006293_human_2511 GAGGACGTGTATGATCTCAT 975 CCTCCGGAGTGTATGGAGGACGTG  995 TATGATCTCATGTACCAGTGC 16 TYRO3_NM_006293_human_3400 TTTTAGGTGAGGGTTGGTAA 976 CCTTGTAATATTCCCTTTTAGGTGA  996 GGGTTGGTAAGGGGTTGGTA 17 TYRO3_NM_006293_human_1895 AGCTGACATCATTGCCTCAA 977 TGTGAAGATGCTGAAAGCTGACAT  997 CATTGCCTCAAGCGACATTGA 18 TYRO3_NM_006293_human_3690 AAATCTCAAAATATCCTAAG 978 TCTGAGCACGCTACCAAATCTCAA  998 AATATCCTAAGACTAACAAAG 19 TYRO3_NM_006293_human_3919 AAGCTGTTAGTTGCTGTTTA 979 GTTAGGGACATTTCCAAGCTGTTA  999 GTTGCTGTTTAAAATAGAAAT 20 TYRO3_NM_006293_human_3384 TCCTTGTAATATTCCCTTTT 980 AGTCACAAAGAGATGTCCTTGTAA 1000 TATTCCCTTTTAGGTGAGGGT Accession: NM_000546 HUGO TP53 gene symbol: SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO:  1 TP53_NM_000546_human_1630 TGTTTGGGAGATGTAAGAAA  81 TTTTACTGTGAGGGATGTTTGGG 101 AGATGTAAGAAATGTTCTTGCA  2 TP53_NM_000546_human_1808 GCATTGTGAGGGTTAATGAA  82 CCTACCTCACAGAGTGCATTGTGA 102 GGGTTAATGAAATAATGTACA  3 TP53_NM_000546_human_2538 TCGATCTCTTATTTTACAAT  83 TATCCCATTTTTATATCGATCTCTT 103 ATTTTACAATAAAACTTTGC  4 TP53_NM_000546_human_1812 TGTGAGGGTTAATGAAATAA  84 CCTCACAGAGTGCATTGTGAGGG 104 TTAATGAAATAATGTACATCTG  5 TP53_NM_000546_human_812 GAGTATTTGGATGACAGAAA  85 GGAAATTTGCGTGTGGAGTATTT 105 GGATGACAGAAACACTTTTCGA  6 TP53_NM_000546_human_1627 GGATGTTTGGGAGATGTAAG  86 GGTTTTTACTGTGAGGGATGTTTG 106 GGAGATGTAAGAAATGTTCTT  7 TP53_NM_000546_human_1646 GAAATGTTCTTGCAGTTAAG  87 GTTTGGGAGATGTAAGAAATGTT 107 CTTGCAGTTAAGGGTTAGTTTA  8 TP53_NM_000546_human_1831 ATGTACATCTGGCCTTGAAA  88 AGGGTTAATGAAATAATGTACAT 108 CTGGCCTTGAAACCACCTTTTA  9 TP53_NM_000546_human_1645 AGAAATGTTCTTGCAGTTAA  89 TGTTTGGGAGATGTAAGAAATGT 109 TCTTGCAGTTAAGGGTTAGTTT 10 TP53_NM_000546_human_2015 GGTGAACCTTAGTACCTAAA  90 GTCTGACAACCTCTTGGTGAACCT 110 TAGTACCTAAAAGGAAATCTC 11 TP53_NM_000546_human_1753 TAACTTCAAGGCCCATATCT  91 CTGTTGAATTTTCTCTAACTTCAA 111 GGCCCATATCTGTGAAATGCT 12 TP53_NM_000546_human_782 CTTATCCGAGTGGAAGGAAA 92 GCCCCTCCTCAGCATCTTATCCGA 112 GTGGAAGGAAATTTGCGTGTG 13 TP53_NM_000546_human_2086 ATGATCTGGATCCACCAAGA   93 CATCTCTTGTATATGATGATCTGG 113 ATCCACCAAGACTTGTTTTAT 14 TP53_NM_000546_human_1744 AATTTTCTCTAACTTCAAGG  94 TGTCCCTCACTGTTGAATTTTCTCT 114 AACTTCAAGGCCCATATCTG 15 TP53_NM_000546_human_2542 TCTCTTATTTTACAATAAAA  95 CCATTTTTATATCGATCTCTTATTT 115 TACAATAAAACTTTGCTGCC 16 TP53_NM_000546_human_2546 TTATTTTACAATAAAACTTT  96 TTTTATATCGATCTCTTATTTTACA 116 ATAAAACTTTGCTGCCACCT 17 TP53_NM_000546_human_1842 GCCTTGAAACCACCTTTTAT  97 AATAATGTACATCTGGCCTTGAAA 117 CCACCTTTTATTACATGGGGT 18 TP53_NM_000546_human_2534 TATATCGATCTCTTATTTTA  98 TTTATATCCCATTTTTATATCGATC 118 TCTTATTTTACAATAAAACT 19 TP53_NM_000546_human_2021 CCTTAGTACCTAAAAGGAAA  99 CAACCTCTTGGTGAACCTTAGTAC 119 CTAAAAGGAAATCTCACCCCA 20 TP53_NM_000546_human_1809 CATTGTGAGGGTTAATGAAA 100 CTACCTCACAGAGTGCATTGTGA 120 GGGTTAATGAAATAATGTACAT 

What is claimed is:
 1. An immune modulator comprising an sdRNA, wherein the sdRNA comprises a guide strand and a passenger strand, wherein the guide strand is about 19-25 nucleotides long, and the passenger strand is about 10-19 nucleotides long, wherein the sdRNA includes a double stranded region and a single stranded region, wherein the single stranded region includes 5-9 phosphorothioate modifications, wherein the sdRNA is chemically modified, including at least one 2′-O-methyl modification or 2′-fluoro modification, wherein the sdRNA targets a sequence selected from SEQ ID NOs: 281-300, and wherein the sdRNA is capable of suppressing expression of PD1.
 2. An immunogenic composition comprising an sdRNA, wherein the sdRNA comprises a guide strand and a passenger strand, wherein the guide strand is about 19-25 nucleotides long, and the passenger strand is about 10-19 nucleotides long, wherein the sdRNA includes a double stranded region and a single stranded region, wherein the single stranded region includes 5-9 phosphorothioate modifications, wherein the sdRNA is chemically modified, including at least one 2′-O-methyl modification or 2′-fluoro modification, wherein the sdRNA targets a sequence selected from SEQ ID NOs: 281-300, wherein the sdRNA is capable of suppressing expression of PD1, and wherein the immunogenic composition further comprises immune cells modified by the sdRNA to suppress expression of PD1.
 3. The immunogenic composition of claim 2, wherein the immune cells within the composition are further modified to suppress expression of a different immune checkpoint gene.
 4. The immunogenic composition of claim 3, wherein said cells are modified to suppress expression of at least one immune checkpoint gene, and at least one anti-apoptosis gene.
 5. The immunogenic composition of claim 2, wherein the immune cells within the composition are further modified to suppress expression of at least one cytokine receptor gene.
 6. The immunogenic composition of claim 3, wherein said cells are modified to suppress expression of at least one immune checkpoint gene and at least one regulator gene.
 7. The immunogenic composition of claim 3, wherein the different immune checkpoint gene is HAVCR2, wherein the composition further comprises an sdRNA capable of suppressing expression of HAVCR2, wherein the sdRNA comprises a guide strand and a passenger strand, wherein the guide strand is about 19-25 nucleotides long, and the passenger strand is about 10-19 nucleotides long, wherein the sdRNA includes a double stranded region and a single stranded region, wherein the single stranded region includes 5-9 phosphorothioate modifications, and wherein the sdRNA is chemically modified, including at least one 2′-O-methyl modification or 2′-fluoro modification.
 8. The immunogenic composition of claim 2, wherein said cells are selected from the group consisting of T-cells, NK-cells, antigen-presenting cells, dendritic cells, stem cells, induced pluripotent stem cells, and/or stem central memory T-cells.
 9. The immunogenic composition of claim 8, wherein said cells are T-cells comprising one or more transgene expressing high affinity T-cell receptors (TCR) and/or chimeric antibody-T-cell receptors (CAR).
 10. The immunogenic composition of claim 7, wherein said cells further comprise one or more sdRNA agent targeting TP53.
 11. The immunogenic composition of claim 2, wherein the sdRNA induces at least 50% inhibition of expression of PD1.
 12. The immunogenic composition of claim 2, wherein the sdRNA comprises at least one hydrophobic modification.
 13. The immunogenic composition of claim 2, wherein the sdRNA is modified to comprises at least one cholesterol molecule.
 14. The immunogenic composition of claim 7, wherein the sdRNA capable of suppressing expression of HAVCR2 targets a sequence selected from SEQ ID NOs: 361-380.
 15. A method of producing the immunogenic composition of claim 2, said method comprising transforming an immune cell with an sdRNA, wherein the sdRNA targets a sequence selected from SEQ ID NOs: 281-300, and wherein the sdRNA is capable of suppressing expression of PD1.
 16. The method of claim 15, wherein the cell further comprises an sdRNA that inhibits expression of HAVCR2, wherein the sdRNA that inhibits expression of HAVCR2 comprises a guide strand and a passenger strand, wherein the guide strand is about 19-25 nucleotides long, and the passenger strand is about 10-19 nucleotides long, wherein the sdRNA includes a double stranded region and a single stranded region, wherein the single stranded region includes 5-9 phosphorothioate modifications, wherein the sdRNA is chemically modified, including at least one 2′-O-methyl modification or 2′-fluoro modification and wherein the sdRNA targets a sequence selected from SEQ ID NOs: 361-380.
 17. The method of claim 15, wherein said cells are selected from the group consisting of T-cells, NK-cells, antigen-presenting cells, dendritic cells, stem cells, induced pluripotent stem cells, and/or stem central memory T-cells.
 18. The method of claim 15, wherein said cells are T-cells comprising a transgene expressing high affinity T-cell receptors (TCR) and/or chimeric antibody-T-cell receptors (CAR). 